CN113447591B - Method for detecting main active ingredients of grassleaf sweelflag rhizome in pharmaceutical preparation - Google Patents
Method for detecting main active ingredients of grassleaf sweelflag rhizome in pharmaceutical preparation Download PDFInfo
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- 239000000825 pharmaceutical preparation Substances 0.000 title claims abstract description 37
- 239000004480 active ingredient Substances 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims description 22
- RKFAZBXYICVSKP-AATRIKPKSA-N alpha-asarone Chemical compound COC1=CC(OC)=C(\C=C\C)C=C1OC RKFAZBXYICVSKP-AATRIKPKSA-N 0.000 claims abstract description 107
- RKFAZBXYICVSKP-WAYWQWQTSA-N beta-asarone Chemical compound COC1=CC(OC)=C(\C=C/C)C=C1OC RKFAZBXYICVSKP-WAYWQWQTSA-N 0.000 claims abstract description 54
- RKFAZBXYICVSKP-UHFFFAOYSA-N beta- asarone Natural products COC1=CC(OC)=C(C=CC)C=C1OC RKFAZBXYICVSKP-UHFFFAOYSA-N 0.000 claims abstract description 53
- 239000000523 sample Substances 0.000 claims abstract description 49
- 238000002360 preparation method Methods 0.000 claims abstract description 40
- 239000012488 sample solution Substances 0.000 claims abstract description 40
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 31
- 239000012085 test solution Substances 0.000 claims abstract description 26
- 239000003960 organic solvent Substances 0.000 claims abstract description 25
- 238000001514 detection method Methods 0.000 claims abstract description 23
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 11
- 239000000706 filtrate Substances 0.000 claims abstract description 8
- 238000001914 filtration Methods 0.000 claims abstract description 8
- 238000001816 cooling Methods 0.000 claims abstract description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 84
- 239000013558 reference substance Substances 0.000 claims description 45
- 239000000243 solution Substances 0.000 claims description 42
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 30
- 239000012088 reference solution Substances 0.000 claims description 21
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical group CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 18
- 239000007864 aqueous solution Substances 0.000 claims description 16
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 15
- 230000014759 maintenance of location Effects 0.000 claims description 12
- 238000010829 isocratic elution Methods 0.000 claims description 10
- 238000012360 testing method Methods 0.000 claims description 10
- 239000008194 pharmaceutical composition Substances 0.000 claims description 9
- 238000004458 analytical method Methods 0.000 claims description 7
- 244000001632 Acorus gramineus Species 0.000 claims description 6
- 235000013073 Acorus gramineus Nutrition 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 238000004090 dissolution Methods 0.000 claims description 6
- 238000010812 external standard method Methods 0.000 claims description 6
- 239000000945 filler Substances 0.000 claims description 4
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 239000003643 water by type Substances 0.000 claims description 2
- 238000003908 quality control method Methods 0.000 abstract description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 15
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 15
- 239000003814 drug Substances 0.000 description 8
- 238000005303 weighing Methods 0.000 description 8
- 238000002347 injection Methods 0.000 description 6
- 239000007924 injection Substances 0.000 description 6
- 238000011084 recovery Methods 0.000 description 5
- 239000007788 liquid Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 208000019901 Anxiety disease Diseases 0.000 description 3
- 230000036506 anxiety Effects 0.000 description 3
- 239000013583 drug formulation Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 239000012528 membrane Substances 0.000 description 3
- 238000005070 sampling Methods 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 241000208340 Araliaceae Species 0.000 description 2
- 206010033557 Palpitations Diseases 0.000 description 2
- 235000005035 Panax pseudoginseng ssp. pseudoginseng Nutrition 0.000 description 2
- 235000003140 Panax quinquefolius Nutrition 0.000 description 2
- 241001080798 Polygala tenuifolia Species 0.000 description 2
- 244000197580 Poria cocos Species 0.000 description 2
- 235000008599 Poria cocos Nutrition 0.000 description 2
- 208000013738 Sleep Initiation and Maintenance disease Diseases 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000008434 ginseng Nutrition 0.000 description 2
- 206010022437 insomnia Diseases 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000009210 therapy by ultrasound Methods 0.000 description 2
- 239000000341 volatile oil Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 244000205574 Acorus calamus Species 0.000 description 1
- 241000544682 Acorus tatarinowii Species 0.000 description 1
- 208000000044 Amnesia Diseases 0.000 description 1
- 208000031091 Amnestic disease Diseases 0.000 description 1
- 241000209524 Araceae Species 0.000 description 1
- 235000011996 Calamus deerratus Nutrition 0.000 description 1
- 206010010305 Confusional state Diseases 0.000 description 1
- 206010012289 Dementia Diseases 0.000 description 1
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- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 241000110847 Kochia Species 0.000 description 1
- 240000006711 Pistacia vera Species 0.000 description 1
- 235000003447 Pistacia vera Nutrition 0.000 description 1
- 208000001431 Psychomotor Agitation Diseases 0.000 description 1
- 206010038743 Restlessness Diseases 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 230000006986 amnesia Effects 0.000 description 1
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- ZPUCINDJVBIVPJ-LJISPDSOSA-N ***e Chemical compound O([C@H]1C[C@@H]2CC[C@@H](N2C)[C@H]1C(=O)OC)C(=O)C1=CC=CC=C1 ZPUCINDJVBIVPJ-LJISPDSOSA-N 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 235000020233 pistachio Nutrition 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 235000007586 terpenes Nutrition 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 230000004580 weight loss Effects 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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N2030/042—Standards
- G01N2030/047—Standards external
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a detection method of main active ingredients of grassleaf sweelflag rhizome in a pharmaceutical preparation, which comprises the following steps: dividing a medicinal preparation sample into a first medicinal preparation sample and/or a second medicinal preparation sample, respectively adding an organic solvent into the first medicinal preparation sample and/or the second medicinal preparation sample, dissolving, performing ultrasonic extraction, cooling, shaking, filtering, respectively obtaining a first sample solution and/or a second sample solution from subsequent filtrate, respectively detecting by adopting a high performance liquid chromatography, and determining main active ingredients of the grassleaf sweelflag rhizome in the first sample solution: the main active ingredients of the grassleaf sweelflag rhizome in the beta-asarone and/or the second test solution are as follows: content of alpha-asarone. The detection method of the main active ingredients of the grassleaf sweelflag rhizome in the medicinal preparation provided by the invention has the advantages of good precision, reproducibility and stability, accuracy and reliability, can truly reflect the quality difference of the grassleaf sweelflag rhizome in the medicinal preparation, and 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 active components of grassleaved sweetflag rhizome in a medicinal preparation, and in particular relates to a detection method of main active components of grassleaved sweetflag rhizome in the medicinal preparation: a method for detecting beta-asarone and alpha-asarone.
Background
The medicinal preparation is first loaded in Tangsun Si far away, the preparation of the prescription for treating urgent need of thousand gold: 'mainly good forgetting prescription', polygala tenuifolia, ginseng (each four parts), poria cocos (two parts) and calamus (one two parts), the upper four ingredients are treated by sieving, and the dagger and the day three are taken orally. The Chinese medicinal composition is used for treating the emotional diseases of heart qi deficiency, restlessness, anxiety, insomnia, depression, and the like, is similar to the depression of Western medicine, and is a basic prescription for improving intelligence, nourishing heart, soothing nerves and stabilizing mind in Chinese medicine. The pharmaceutical preparation is used for treating depression, anxiety and dementia in modern clinic, and the clinical manifestations of the pharmaceutical preparation are usually mental confusion, absentmindedness, anxiety, amnesia, insomnia, palpitation, severe palpitation and the like.
The medicine preparation is brown yellow powder and consists of 4 traditional Chinese medicines of ginseng, polygala tenuifolia, grassleaved sweetflag rhizome and poria cocos. Rhizoma Acori Graminei is dried rhizome of Acorus tatarinowii Acorus tatarinowii Schott of Araceae, and contains volatile oil, organic acid, terpenes, flavone, etc., wherein the main active ingredient is volatile oil, and has effects of eliminating phlegm, inducing resuscitation, eliminating dampness, stimulating appetite, refreshing mind, and improving intelligence, and mainly contains beta-asarone, alpha-asarone, etc. Because of the lack of a quantitative detection method for main active ingredients of the grassleaf sweelflag rhizome in the medicinal preparation at present, a corresponding method is necessary to be established, the quality control of the grassleaf sweelflag rhizome is carried out, and the quality control system of the medicinal preparation is perfected.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, an object of the present invention is to provide a method for detecting a main active ingredient of rhizoma Acori Graminei in a pharmaceutical preparation, which is used for solving the problem that the main active ingredient of rhizoma Acori Graminei in the pharmaceutical preparation is lacking in the prior art: and the content determination method of the beta-asarone and the alpha-asarone.
To achieve the above and other related objects, a first aspect of the present invention provides a method for detecting a major active ingredient of grassleaf sweelflag rhizome in a pharmaceutical preparation, comprising: dividing a medicinal preparation sample into a first medicinal preparation sample and/or a second medicinal preparation sample, respectively adding an organic solvent into the first medicinal preparation sample and/or the second medicinal preparation sample, dissolving, performing ultrasonic extraction, cooling, shaking, filtering, respectively obtaining a first sample solution and/or a second sample solution from subsequent filtrate, respectively detecting by adopting a high performance liquid chromatography, and determining main active ingredients of the grassleaf sweelflag rhizome in the first sample solution: the main active ingredients of the grassleaf sweelflag rhizome in the beta-asarone and/or the second test solution are as follows: content of alpha-asarone.
Preferably, the CAS number of the beta-asarone is 5273-86-9, and the CAS number of the alpha-asarone is 2883-98-9.
Preferably, the ratio of the mass of the first pharmaceutical preparation sample added to the volume of the organic solvent added is 0.25-8:25, g/mL. Preferably, the ratio of the mass of the first pharmaceutical formulation sample addition to the volume of organic solvent addition is 0.5:25, g/mL.
Preferably, the ratio of the mass of the second pharmaceutical preparation sample added to the volume of the organic solvent added is 0.25-12:25, g/mL. Preferably, the ratio of the mass of the second pharmaceutical formulation sample addition to the volume of the organic solvent addition is 4:25, g/mL.
Preferably, the organic solvent is selected from one of methanol, 70% methanol, 50% methanol, ethanol or ethyl acetate. Preferably, the organic solvent is methanol.
The 70% methanol and the 50% methanol are respectively 70% and 50% methanol aqueous solutions by volume percent.
Preferably, the ultrasonic extraction time is 30-60 min. Preferably, the ultrasonic extraction time is 30min.
Preferably, the power of ultrasonic extraction is 300-400W, and the frequency of ultrasonic extraction is 50-60 kHz. Preferably, the power of the ultrasonic extraction is 350W, and the frequency of the ultrasonic extraction is 53kHz.
Preferably, the first pharmaceutical formulation sample and/or the second pharmaceutical formulation sample is precisely weighed after being added to the organic solvent.
Preferably, the cooling is followed by re-weighing and the weight loss is compensated with an organic solvent.
Preferably, the filtering means: taking a supernatant filtering membrane of the uniformly-shaken solution, and discarding the primary filtrate to obtain the subsequent filtrate.
More preferably, the filter is a 0.22 μm filter.
Preferably, the high performance liquid chromatography performs detection respectively, and includes the following steps:
1) Preparing a first reference substance solution and/or a second reference substance solution: adding organic solvent into the beta-asarone reference substance and/or the alpha-asarone reference substance respectively to dissolve and fix the volume to prepare a first reference substance solution and/or a second reference substance solution;
2) Sample detection: and respectively detecting the first test solution and/or the second test solution, the first reference solution and/or the second reference solution by adopting a high performance liquid chromatography method, comparing the retention time of the first test solution and the first reference solution and/or the retention time of the second test solution and the second reference solution, performing qualitative determination, and quantifying by adopting an external standard method to determine the content of beta-asarone in the first test solution and/or alpha-asarone in the second test solution.
Preferably, in step 1), the organic solvent is selected from one of methanol, 70% methanol, 50% methanol, ethanol or ethyl acetate. More preferably, the organic solvent is methanol.
The 70% methanol and the 50% methanol are respectively 70% and 50% methanol aqueous solutions by volume percent.
Preferably, in the step 1), the content of the beta-asarone in the first reference substance solution ranges from 10.125 to 324 mug/mL.
Preferably, in the step 1), the content range of the alpha-asarone in the second reference substance solution is 1.60-51.32 mug/mL.
Preferably, in step 1), the first reference substance solution and/or the second reference substance solution are diluted stepwise.
Preferably, in step 2), the high performance liquid chromatography method uses a Diode Array Detector (DAD).
Preferably, in step 2), in the high performance liquid chromatography, a chromatographic column is used as C 18 Chromatographic column (4.6X250 mm,5 μm)The filler is octadecylsilane chemically bonded silica gel.
More preferably, in the high performance liquid chromatography, the chromatographic column is selected from Agilent ZORBAX SB-C 18 Chromatographic columns (4.6X250 mm,5 μm), agilent Eclipse Plus C 18 Chromatographic columns (4.6X105 mm,5 μm) or WatersC 18 One of the columns (4.6X250 mm,5 μm), preferably AgilentZORBAX SB-C 18 A chromatographic column.
Preferably, in the step 2), the high performance liquid chromatography uses a detector wavelength of 250-260 nm, preferably 254nm.
Preferably, in step 2), the column temperature is 25 to 40 ℃ in the high performance liquid chromatography. More preferably, the column temperature is 35 ℃.
Preferably, in the step 2), the flow rate of the mobile phase is 1.3-1.7 ml/min in the high performance liquid chromatography. More preferably, the mobile phase is at a flow rate of 1.5ml/min.
Preferably, in the step 2), the sample injection amount is 5 to 20 μl in the high performance liquid chromatography. More preferably, the sample injection amount is 10 μl.
Preferably, in the step 2), in the high performance liquid chromatography, the mobile phase is acetonitrile-0.09-0.11% phosphoric acid aqueous solution, wherein the phase A is acetonitrile and the phase B is 0.09-0.11% phosphoric acid aqueous solution; the analysis time is 25-60min; isocratic elution.
More preferably, in the high performance liquid chromatography, the mobile phase is acetonitrile-0.1% phosphoric acid aqueous solution, wherein the A phase is acetonitrile and the B phase is 0.1% phosphoric acid aqueous solution; the analysis time is 30min; isocratic elution.
More preferably, in the isocratic elution, phase a: the volume ratio of the phase B is 35-45: 55-65.
Further preferably, in the isocratic elution, phase a: the volume ratio of the phase B is 40:60.
the 0.09-0.11% phosphoric acid aqueous solution is 0.09-0.11% phosphoric acid aqueous solution by volume percent. The 0.1% phosphoric acid aqueous solution is a 0.1% phosphoric acid aqueous solution by volume. Preferably, in step 2), the external standard method comprises the following steps:
a) Preparing a series of first reference substance solutions and/or second reference substance solutions with different concentrations according to the step 1), respectively performing High Performance Liquid Chromatography (HPLC) detection to obtain linear relation between chromatographic peak areas of the beta-asarone and/or alpha-asarone and the concentrations of the corresponding beta-asarone and/or alpha-asarone, drawing a corresponding standard working curve, and calculating to obtain a regression equation of the standard working curve of the beta-asarone and/or alpha-asarone;
b) And C, detecting the first sample solution and/or the second sample solution by High Performance Liquid Chromatography (HPLC), substituting the chromatographic peak area of the obtained beta-asarone and/or alpha-asarone into a regression equation of the standard working curve of the corresponding beta-asarone and/or alpha-asarone in the step A), and calculating to obtain the content of the beta-asarone in the first sample solution and/or the content of the alpha-asarone in the second sample solution.
More preferably, the standard working curve is plotted on the ordinate (Y-axis) against the chromatographic peak area of β -asarone and/or α -asarone, and the concentration of the corresponding β -asarone and/or α -asarone is plotted on the abscissa (X-axis).
The second aspect of the invention provides an application of a detection method of main active ingredients of grassleaved sweetflag in a pharmaceutical preparation in quality detection of grassleaved sweetflag in the pharmaceutical preparation.
The medicinal preparation is a medicinal preparation containing rhizoma acori graminei, and specifically comprises, but is not limited to, pistachio powder and the like.
The third aspect of the invention provides a quality detection method of grassleaf sweelflag rhizome in a pharmaceutical preparation, wherein the grassleaf sweelflag rhizome is treated by beta-asarone (C 12 H 16 O 3 ) Is more than or equal to 0.06 percent based on alpha-asarone (C) 12 H 16 O 3 ) The content is more than or equal to 0.04 percent.
As described above, the method for detecting the active ingredients of the grassleaf sweelflag rhizome in the pharmaceutical preparation provided by the invention adopts pretreatment with optimized conditions and an instrument detection method, and is used for detecting the main active ingredients of the grassleaf sweelflag rhizome in the pharmaceutical preparation: and the beta-asarone and the alpha-asarone are accurately, quantitatively and qualitatively detected. The method comprises the steps of preparing two samples, namely preparing two sample solutions with different feed liquid ratios, and carrying out 2 liquid chromatography analysis and determination to respectively determine the contents of beta-asarone and alpha-asarone in the pharmaceutical preparation. The method has good precision, reproducibility and stability, accurate and reliable result, can truly reflect the quality difference of rhizoma Acori Graminei in the pharmaceutical preparation, ensures the stability of the production process and quality between batches, and comprehensively improves the quality control system of the pharmaceutical preparation.
Drawings
FIG. 1 shows a beta-asarone specific chromatogram of example 2 of the present invention
FIG. 2 shows a specific chromatogram of α -asarone of example 2 of the present invention
FIG. 3 shows a graph of concentration versus peak area for the beta-asarone of example 3 of the present invention
FIG. 4 shows a graph of concentration versus peak area for the alpha-asarone of example 3 of the present invention
Detailed Description
The invention is further illustrated below in connection with specific examples, which are to be understood as being illustrative of the invention and not limiting the scope of the invention.
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention.
The reagents and instrumentation used in the following examples were as follows:
1. reagent(s)
Pharmaceutical formulations (Shanghai and yellow pharmaceutical Co., ltd.); beta-asarone reference (purity 99.3%, chinese food and drug inspection institute, lot 112018-201802); alpha-asarone reference (purity is more than or equal to 98 percent, shanghai Shiadad standard technical service Co., ltd., batch number is 4940); acetonitrile (chromatographic purity, lot number JA089830, merck); phosphoric acid (chromatographic purity, national drug); methanol, ethanol, ethyl acetate (analytically pure, chinese medicine); deionized water (self-made by water purifier).
2. Instrument for measuring and controlling the intensity of light
SK7200H ultrasonic cleaner (Shanghai Kochia ultrasonic instruments Co., ltd.); ME204E/02 type electronic balance (Metrele Tolyduo instruments (Shanghai) Co., ltd.); agilent 1260 high performance liquid chromatograph (Agilent technologies, inc.) and Agilent 1260II high performance liquid chromatograph (Agilent technologies, inc.).
The content of the main active ingredients of the grassleaf sweelflag rhizome in the pharmaceutical preparation comprises the following determination process.
1. Preparation of test solutions
The preparation method comprises the steps of taking a preparation sample of the medicine, dividing the preparation sample into a first preparation sample and/or a second preparation sample, respectively adding an organic solvent for dissolution, sealing, performing ultrasonic extraction (300-400 w, 50-60 kHz) for 30-60 minutes, cooling, shaking uniformly, taking supernatant, filtering, and taking subsequent filtrate to obtain a first sample solution and/or a second sample solution respectively. Wherein the ratio of the mass of the first pharmaceutical preparation sample added to the volume of the organic solvent added is 0.25-8:25, g/mL. The ratio of the mass of the second pharmaceutical preparation sample added to the volume of the organic solvent added is 0.25-12:25, g/mL. The organic solvent is selected from one of methanol, 70% methanol, 50% methanol, ethanol or ethyl acetate.
2. Preparation of control solution
Adding the beta-asarone reference substance into an organic solvent for dissolution and volume fixing, and preparing a first reference substance solution, wherein the content range of the first reference substance solution is 10.125-324 mug/mL. Adding the alpha-asarone reference substance into an organic solvent for dissolution and volume fixing, and preparing a second reference substance solution, wherein the content range of the second reference substance solution is 1.60-51.32 mug/mL. The organic solvent is selected from one of methanol, 70% methanol, 50% methanol, ethanol or ethyl acetate.
3. Measurement
And (3) respectively detecting the first test solution and/or the second test solution and the first reference solution and/or the second reference solution by adopting a high performance liquid chromatography, comparing the retention time of the first test solution and the first reference solution and/or the retention time of the second test solution and the second reference solution, carrying out qualitative determination by adopting an external standard method, namely respectively detecting a series of first reference solutions and/or second reference solutions with different concentrations by adopting a High Performance Liquid Chromatography (HPLC), obtaining a linear relation between the chromatographic peak area of the beta-asarone and/or the alpha-asarone and the concentration of the corresponding beta-asarone and/or the alpha-asarone, drawing a corresponding standard working curve, and calculating to obtain a regression equation of the standard working curve of the beta-asarone and/or the alpha-asarone. And then carrying out High Performance Liquid Chromatography (HPLC) detection on the first sample solution and/or the second sample solution, substituting the chromatographic peak area of the obtained beta-asarone and/or alpha-asarone into a regression equation of a standard working curve of the corresponding beta-asarone and/or alpha-asarone, and calculating to obtain the content of the beta-asarone in the first sample solution and/or the content of the alpha-asarone in the second sample solution.
Wherein, the high performance liquid chromatography comprises the following detection conditions:
the detector is a Diode Array Detector (DAD); the spectrum column is C 18 A chromatographic column (4.6X250 mm,5 μm), wherein the filler in the chromatographic column is octadecylsilane chemically bonded silica; the wavelength of the detector is 250-260 nm; the column temperature is 25-40 ℃; the flow rate is 1.3-1.7 ml/min; the sample injection amount is 5-20 mu L; the mobile phase is acetonitrile-0.09-0.11% phosphoric acid aqueous solution, wherein, the A phase is acetonitrile, and the B phase is 0.09-0.11% phosphoric acid aqueous solution; the analysis time is 25-60min; isocratic elution. In isocratic elution, phase a: the volume ratio of the phase B is 35-45: 55-65.
Example 1
1. Preparation of test solutions
The drug formulation sample is divided into a first drug formulation sample and a second drug formulation sample.
Taking 0.50g of a first pharmaceutical preparation sample, precisely weighing, placing into a conical flask with a plug, precisely adding 25mL of methanol, sealing, performing ultrasonic treatment (350 w,53 kHz) for 30 minutes, cooling, shaking uniformly, taking supernatant, and filtering with a 0.22 mu m filter membrane, and taking subsequent filtrate to obtain a first sample solution No. 1.
Taking 4.00g of a second pharmaceutical preparation sample, precisely weighing, placing into a conical flask with a plug, precisely adding 25mL of methanol, sealing, performing ultrasonic treatment (350 w,53 kHz) for 30 minutes, shaking uniformly, taking supernatant, and filtering with a 0.22 mu m filter membrane to obtain a subsequent filtrate to obtain a second sample solution No. 1.
2. Preparation of control solution
Precisely weighing the beta-asarone reference substance, adding methanol for dissolution and fixing volume to prepare a first reference substance solution No. 1, wherein the content range of the first reference substance solution No. 1 is 10.125-324 mug/mL. Precisely weighing the alpha-asarone reference substance, adding methanol for dissolution and fixing volume to prepare a second reference substance solution No. 1, wherein the content range of the second reference substance solution No. 1 is 1.60-51.32 mug/mL.
3. Measurement
And (3) respectively detecting the first sample solution 1# and the first reference substance solution 1# by adopting a high performance liquid chromatography, comparing the retention time of the first sample solution 1# and the retention time of the first reference substance solution 1#, and quantifying by adopting an external standard method, namely respectively detecting a series of first reference substance solutions 1# with different concentrations by adopting a High Performance Liquid Chromatography (HPLC) method to obtain the linear relation between the chromatographic peak area of the beta-asarone and the corresponding concentration of the beta-asarone, drawing a corresponding standard working curve, and calculating to obtain a regression equation of the standard working curve of the beta-asarone. And then carrying out High Performance Liquid Chromatography (HPLC) detection on the first sample solution 1# and substituting the chromatographic peak area of the obtained beta-asarone into a regression equation of a standard working curve of the corresponding beta-asarone to calculate and obtain the content of the beta-asarone in the first sample solution 1#.
And (3) respectively detecting the second sample solution 1# and the second reference substance solution 1# by adopting a high performance liquid chromatography, comparing the retention time of the second sample solution 1# and the retention time of the second reference substance solution 1# for qualitative determination, and quantifying by adopting an external standard method, namely respectively detecting a series of second reference substance solutions 1# with different concentrations by adopting a High Performance Liquid Chromatography (HPLC) to obtain the linear relation between the chromatographic peak area of the alpha-asarone and the concentration of the corresponding alpha-asarone, drawing a corresponding standard working curve, and calculating to obtain a regression equation of the standard working curve of the alpha-asarone. And then carrying out High Performance Liquid Chromatography (HPLC) detection on the second sample solution 1# and substituting the chromatographic peak area of the obtained alpha-asarone into a regression equation of a standard working curve of the corresponding alpha-asarone to calculate and obtain the content of the alpha-asarone in the second sample solution 1#.
Wherein, the high performance liquid chromatography comprises the following detection conditions:
the detector is a Diode Array Detector (DAD); the spectrum column is Agilent Eclipse Plus C 18 Chromatographic column (4.6X250 mm,5 μm; inner diameter X column length, filler particle size); the detector wavelength is 254nm; the column temperature is 35 ℃; the flow rate is 1.5ml/min; the sample injection amount is 10 mu L; the mobile phase is acetonitrile-0.1% phosphoric acid aqueous solution, wherein the A phase is acetonitrile, and the B phase is 0.1% phosphoric acid aqueous solution; the analysis time is 30min; isocratic elution. In isocratic elution, phase a: the volume ratio of the phase B is 40:60.
example 2
A sample of the pharmaceutical formulation of lot 20200514 was taken and the first and second test solutions were prepared as in step 1 of example 1. Taking a negative sample of the pharmaceutical preparation from which the grassleaf sweelflag rhizome is removed, and preparing a first negative test solution and a second negative test solution by adopting the step 1 in the embodiment 1.
Meanwhile, a first reference solution and a second reference solution were prepared as in step 2 of example 1, wherein the content of β -asarone in the first reference solution was 10.125 μg/mL, and the content of α -asarone in the second reference solution was 12.83 μg/mL.
The first sample solution, the first reference solution and the first negative sample solution were measured according to step 3 in example 1, and the retention time was compared and the results were confirmed as shown in fig. 1. The second sample solution, the second negative sample solution and the second control solution were measured according to step 3 in example 1, and the retention time was compared and the results were confirmed as shown in fig. 2. As shown in figures 1 and 2, the determination shows that the negative sample has no interference to the test sample and has good specificity.
Example 3
The detection method of the main active ingredients of the grassleaf sweelflag rhizome in the pharmaceutical preparation is subjected to methodological verification, and the performance index results are as follows.
1. Linear relation of detection method
Respectively precisely weighing appropriate amounts of beta-asarone and alpha-asarone reference substances, and adding methanol according to step 2 in example 1 to obtain a series of first reference substance solutions and second reference substance solutions with different concentrations. According to the chromatographic conditions of step 3 in example 1, 10 μl of the first reference solution and the second reference solution are precisely sucked and injected into a high performance liquid chromatograph, the concentrations of the beta-asarone and the alpha-asarone are plotted with the abscissa, the chromatographic peak areas of the beta-asarone and the alpha-asarone are plotted with the ordinate, and the standard regression equation, the correlation coefficient and the linear range of 2 components are measured and calculated, and specific results are shown in tables 1 and figures 3-4. As can be seen from table 1, fig. 3-4, the linear equation for β -asarone is: y=20.399068 x+22.17463, r= 0.99996, indicating a good linear relationship in the linear range 0.10125-3.24 μg; the linear equation for α -asarone is: y=27.448511+2.58433, r=0.99999, indicating 0.0160375 to 0.5132 μg.
TABLE 1
2. Stability of
Samples of the pharmaceutical preparation (lot number: 20200514) were taken, a first test solution and a second test solution were prepared according to step 1 of example 1, sample analysis was performed at 0h, 2h, 4h, 6h, 12h, 18h, 24h, 36h, respectively, according to the chromatographic conditions of step 3 of example 1, peak areas were determined, and the content of β -asarone and α -asarone was calculated, and specific stability results are shown in table 2. The results showed that the RSD for the beta-asarone content was 0.17% and the RSD for the alpha-asarone content was 0.41%. The RSD of the content of the first test sample solution and the second test sample solution is less than 0.5%, and the RSD is basically stable within 36 hours, which indicates that the method has good stability.
TABLE 2
3. Precision of
3.1 precision of instrument
Taking appropriate amounts of beta-asarone and alpha-asarone reference substances, precisely weighing, adding methanol to prepare a first reference substance solution and a second reference substance solution with certain concentrations according to the step 2 in the embodiment 1, continuously sampling for 6 times according to the chromatographic conditions of the step 3 in the embodiment 1, and measuring peak areas, wherein the results are shown in Table 3. As shown in Table 3, the RSD of the 6 times of injection peak areas of the beta-asarone is 0.15%, the RSD of the 6 times of injection peak areas of the alpha-asarone is 0.12%, and the RSD of the chromatographic peak areas of the 2 components is less than 0.2%, which indicates that the instrument precision is good.
TABLE 3 Table 3
3.2 repeatability
6 parts of the same batch of pharmaceutical preparation samples (batch number: 20200514) were taken, 6 parts of a first test sample solution and 6 parts of a second test sample solution were prepared and obtained according to the step 1 in example 1, and the sample was introduced and analyzed respectively according to the chromatographic conditions of the step 3 in example 1, and the peak area was measured, and the specific repeatability results are shown in Table 4. The results showed that the chromatographic peak areas RSD of the 2 components were all less than 0.4%, indicating good reproducibility of the method.
TABLE 4 Table 4
3.3 intermediate precision
The same batch of samples of the pharmaceutical preparation was taken by different laboratory workers, 2 parts of the first test substance solution and 2 parts of the second test substance solution were prepared in step 1 of example 1, and the results were shown in Table 5 according to the chromatographic conditions of step 3 of example 1. RSD of 2 component contents in samples measured by different experimenters and different instruments is less than 5%, which indicates that the intermediate precision is good.
TABLE 5
4. Recovery rate of sample addition
Precisely weighing 9 parts of pharmaceutical preparation (batch No. 20200514) with known concentration 0.25g, and adding beta-asarone reference substance solution at three levels of 50%, 100% and 150% of the content, wherein each level is divided into three parts in parallel; 9 parts of a pharmaceutical preparation (batch No. 20200514) of known concentration (2.00 g) are precisely weighed, and alpha-asarone reference substance solutions are respectively added at three levels of 50%, 100% and 150% of the content, and each level is divided into three parts in parallel. The first test sample solution and the second test sample solution were prepared in step 1 of example 1, and analyzed under the chromatographic conditions of step 3 of example 1, and the results are shown in Table 6. As shown in Table 6, the recovery rate of the beta-asarone is between 90 and 108 percent, the recovery rate of the alpha-asarone is between 80 and 115 percent, and meets the recovery rate limit specified in the 'Chinese pharmacopoeia' of 2020 edition (four parts), which proves that the recovery rate of the method is good.
TABLE 6
Example 4
The first test solution and the second test solution were prepared in step 1 of example 1 for 6 different sampling point samples of pharmaceutical formulation lot S210301R01, and analyzed according to the chromatographic conditions of step 3 of example 1. The content measurement results are shown in Table 7. As can be seen from Table 7, the RSD of the contents of the beta-asarone and the alpha-asarone in the samples of 6 different sampling points of the S210301R01 batch of the pharmaceutical preparation is less than 5%, which indicates that the S210301R01 batch of the Happy powder satisfies the requirement of mixing uniformity.
TABLE 7 determination of content uniformity in samples of pharmaceutical formulations
In conclusion, the detection method of the main active ingredients of the grassleaf sweelflag rhizome in the medicinal preparation provided by the invention has the advantages of good precision, reproducibility and stability, accuracy and reliability, can truly reflect the quality difference of the grassleaf sweelflag rhizome in the medicinal preparation, and 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 above embodiments are merely illustrative of the principles of the present invention and its effectiveness, and are not intended to limit the invention. Modifications and variations may be made to the above-described embodiments by those skilled in the art without departing from the spirit and scope of the invention. Accordingly, it is intended that all equivalent modifications and variations of the invention be covered by the claims, which are within the ordinary skill of the art, be within the spirit and scope of the present disclosure.
Claims (4)
1. A method for detecting main active components of rhizoma Acori Graminei in pharmaceutical preparation comprises: dividing a medicinal preparation sample into a first medicinal preparation sample and a second medicinal preparation sample, respectively adding an organic solvent into the first medicinal preparation sample and the second medicinal preparation sample for dissolution, performing ultrasonic extraction, cooling, shaking uniformly, filtering, taking the subsequent filtrate to respectively obtain a first sample solution and a second sample solution, respectively detecting by adopting a high performance liquid chromatography, and determining main active ingredients of the grassleaf sweelflag rhizome in the first sample solution: the main active components of the grassleaf sweelflag rhizome in the solution of the beta-asarone and the second test sample are as follows: content of alpha-asarone;
the medicinal preparation is yippee powder;
the high performance liquid chromatography method respectively detects the components and comprises the following steps:
1) Preparing a first reference substance solution and a second reference substance solution: adding organic solvent into the beta-asarone reference substance and the alpha-asarone reference substance respectively to dissolve and fix the volume to prepare a first reference substance solution and a second reference substance solution;
2) Sample detection: detecting the first and second test solutions, the first and second reference solutions respectively by high performance liquid chromatography, comparing the retention time of the first test solution and the first reference solution with the retention time of the second test solution and the second reference solution, performing qualitative determination, and determining the content of beta-asarone in the first test solution and alpha-asarone in the second test solution by external standard method;
the ratio of the added mass of the first pharmaceutical preparation sample to the added volume of the organic solvent is 0.5:25, g/mL;
the ratio of the mass of the second pharmaceutical formulation sample addition to the volume of the organic solvent addition was 4:25, g/mL;
the organic solvent is methanol;
in the step 2), in the high performance liquid chromatography, the detector is a diode array detector; the chromatographic column is selected from Agilent ZORBAX SB-C 18 Chromatographic column, agilent Eclipse Plus C 18 Chromatographic column or Waters XBidge cube C 18 One of the chromatographic columns has the specification of 4.6X1250 mm and 5 mu m, and the filler in the chromatographic column is octadecylsilane chemically bonded silica gel; the detector wavelength is 254nm; the mobile phase is acetonitrile-0.1% phosphoric acid aqueous solution, wherein the A phase is acetonitrile, and the B phase is 0.1% phosphoric acid aqueous solution; the analysis time is 30min; isocratic elution;
in the isocratic elution, phase a: the volume ratio of the phase B is 40:60;
in the step 2), in the high performance liquid chromatography, the column temperature is 35 ℃; the flow rate is 1.5ml/min; the sample loading was 10. Mu.L.
2. The method for detecting the main active ingredients of grassleaved sweetflag rhizome in a pharmaceutical preparation according to claim 1, wherein the ultrasonic extraction time is 30-60 min.
3. The method for detecting the main active ingredients of grassleaf sweelflag rhizome in a pharmaceutical preparation according to claim 1, wherein in step 1), the content of beta-asarone in the first reference solution is 10.125-324 μg/mL; the content range of the alpha-asarone in the second reference substance solution is 1.60-51.32 mug/mL.
4. Use of a method for detecting the main active ingredient of grassleaf sweelflag rhizome in a pharmaceutical preparation according to any one of claims 1-3 for quality detection of grassleaf sweelflag rhizome in a pharmaceutical preparation.
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Non-Patent Citations (5)
| Title |
|---|
| 巴寅颖等.开心散血清HPLC特征图谱研究.《北京中医药大学学报》.2011,第34卷(第6期),409-412,416. * |
| 汤佳佳等.HPLC 法测定复方麝香注射液中细辛醚.《中成药》.2012,第34卷(第2期),289-292. * |
| 许伟等.一测多评法同时测定宁神定志丸中8种成分.《中成药》.2019,第41卷(第11期),2586-2590. * |
| 陈日来等.小儿遗尿分散片的制剂处方筛选与α－细辛醚、β － 细辛醚含量测定.《中国药业》.2011,第20卷(第21期),40-42. * |
| 陈颖等.经典名方开心散不同温度高压蒸汽灭菌条件下9种指标成分含量变化研究3.中草药.2021,第52卷(第4期),976-981. * |
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