CN1544920A - Method for determining content of Lamiophlomis rotata in Lamiophlomis rotata preparation and measurement standard thereof - Google Patents
Method for determining content of Lamiophlomis rotata in Lamiophlomis rotata preparation and measurement standard thereof Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 52
- 238000000034 method Methods 0.000 title claims abstract description 24
- 241001191006 Phlomoides rotata Species 0.000 title claims description 22
- 238000005259 measurement Methods 0.000 title claims description 11
- 229930003944 flavone Natural products 0.000 claims abstract description 22
- 235000011949 flavones Nutrition 0.000 claims abstract description 22
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 claims abstract description 21
- 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 claims abstract description 21
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 claims abstract description 21
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 claims abstract description 21
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 claims abstract description 21
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 claims abstract description 21
- 235000005493 rutin Nutrition 0.000 claims abstract description 21
- 229960004555 rutoside Drugs 0.000 claims abstract description 21
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 claims abstract description 21
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 claims abstract description 20
- 150000002212 flavone derivatives Chemical class 0.000 claims abstract description 19
- IQPNAANSBPBGFQ-UHFFFAOYSA-N luteolin Chemical compound C=1C(O)=CC(O)=C(C(C=2)=O)C=1OC=2C1=CC=C(O)C(O)=C1 IQPNAANSBPBGFQ-UHFFFAOYSA-N 0.000 claims abstract description 19
- LRDGATPGVJTWLJ-UHFFFAOYSA-N luteolin Natural products OC1=CC(O)=CC(C=2OC3=CC(O)=CC(O)=C3C(=O)C=2)=C1 LRDGATPGVJTWLJ-UHFFFAOYSA-N 0.000 claims abstract description 19
- 235000009498 luteolin Nutrition 0.000 claims abstract description 19
- 239000000243 solution Substances 0.000 claims description 71
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 39
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 32
- 239000002775 capsule Substances 0.000 claims description 30
- 239000013558 reference substance Substances 0.000 claims description 27
- 238000012360 testing method Methods 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 238000003556 assay Methods 0.000 claims description 20
- 230000007062 hydrolysis Effects 0.000 claims description 19
- 238000006460 hydrolysis reaction Methods 0.000 claims description 19
- FUKUFMFMCZIRNT-UHFFFAOYSA-N hydron;methanol;chloride Chemical compound Cl.OC FUKUFMFMCZIRNT-UHFFFAOYSA-N 0.000 claims description 16
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 claims description 16
- 238000001514 detection method Methods 0.000 claims description 13
- 238000002835 absorbance Methods 0.000 claims description 12
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 239000000203 mixture Substances 0.000 claims description 9
- 238000002798 spectrophotometry method Methods 0.000 claims description 9
- 238000005303 weighing Methods 0.000 claims description 9
- BNGXYYYYKUGPPF-UHFFFAOYSA-M (3-methylphenyl)methyl-triphenylphosphanium;chloride Chemical compound [Cl-].CC1=CC=CC(C[P+](C=2C=CC=CC=2)(C=2C=CC=CC=2)C=2C=CC=CC=2)=C1 BNGXYYYYKUGPPF-UHFFFAOYSA-M 0.000 claims description 8
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 8
- 229930003935 flavonoid Natural products 0.000 claims description 8
- 150000002215 flavonoids Chemical class 0.000 claims description 8
- 235000017173 flavonoids Nutrition 0.000 claims description 8
- 208000030208 low-grade fever Diseases 0.000 claims description 8
- 238000002156 mixing Methods 0.000 claims description 8
- 238000007254 oxidation reaction Methods 0.000 claims description 8
- 239000011734 sodium Substances 0.000 claims description 8
- 229910052708 sodium Inorganic materials 0.000 claims description 8
- 235000010288 sodium nitrite Nutrition 0.000 claims description 8
- 239000000126 substance Substances 0.000 claims description 8
- 239000012085 test solution Substances 0.000 claims description 8
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 7
- DLYUQMMRRRQYAE-UHFFFAOYSA-N tetraphosphorus decaoxide Chemical compound O1P(O2)(=O)OP3(=O)OP1(=O)OP2(=O)O3 DLYUQMMRRRQYAE-UHFFFAOYSA-N 0.000 claims description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 5
- 239000000706 filtrate Substances 0.000 claims description 5
- 238000004811 liquid chromatography Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 4
- 238000010438 heat treatment Methods 0.000 claims description 4
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 4
- 239000000377 silicon dioxide Substances 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000000443 aerosol Substances 0.000 claims description 2
- 239000010408 film Substances 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 239000003826 tablet Substances 0.000 claims description 2
- 238000009472 formulation Methods 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 18
- 229940079593 drug Drugs 0.000 abstract description 12
- 239000002021 butanolic extract Substances 0.000 description 25
- 239000002904 solvent Substances 0.000 description 9
- 238000011084 recovery Methods 0.000 description 6
- 239000000284 extract Substances 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 2
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 2
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 description 2
- 239000012490 blank solution Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- YWEUIGNSBFLMFL-UHFFFAOYSA-N diphosphonate Chemical compound O=P(=O)OP(=O)=O YWEUIGNSBFLMFL-UHFFFAOYSA-N 0.000 description 2
- -1 flavone compound Chemical class 0.000 description 2
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000013112 stability test Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- 235000008100 Ginkgo biloba Nutrition 0.000 description 1
- 244000194101 Ginkgo biloba Species 0.000 description 1
- 239000009429 Ginkgo biloba extract Substances 0.000 description 1
- 229930194248 Licoflavone Natural products 0.000 description 1
- MEHHCBRCXIDGKZ-UHFFFAOYSA-N Licoflavone C Natural products CC(C)=CCC1=C(O)C=C(O)C(C(C=2)=O)=C1OC=2C1=CC=C(O)C=C1 MEHHCBRCXIDGKZ-UHFFFAOYSA-N 0.000 description 1
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 description 1
- 241001128140 Reseda Species 0.000 description 1
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000036592 analgesia Effects 0.000 description 1
- 230000010165 autogamy Effects 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000004737 colorimetric analysis Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 150000002213 flavones Chemical class 0.000 description 1
- 229940068052 ginkgo biloba extract Drugs 0.000 description 1
- 235000020686 ginkgo biloba extract Nutrition 0.000 description 1
- 230000023597 hemostasis Effects 0.000 description 1
- 230000002439 hemostatic effect Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000036407 pain Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000005875 quercetin Nutrition 0.000 description 1
- 229960001285 quercetin Drugs 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000004506 ultrasonic cleaning Methods 0.000 description 1
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a method to determine the content of single-taste in single-taste preparation and determination standard, belonging to Chinese medicinal field. It is implemented by determining the content of total flavone and luteolin in single-taste preparation, where the detecting standard is that the total flavone in every 0.3g of single-taste preparation (equivalent to 1 g of crude drug) shouldn't be less than 26.0mg according to rutin (C27H30O16), and the luteolin is 0.8000mg-1.8105mg, i.e. the luteolin is 0.2666%-0.6035%; it makes the quality of the single-taste preparation more stable and reliable.
Description
Technical field
The present invention relates to a kind of medicine detection method, particularly relate to a kind of method of lamiophlomis rotata content in the common lamiophlomis root preparation and standard of mensuration thereof measured.
Background technology
Tibet medicine lamivphlomis root has promoting blood circulation and removing blood stasisly as a kind of, and the medicinal material of pain easing and hemostasis function is usually used in preparing the medicine of hemostatic analgesia.The content that accurately detects lamiophlomis rotata in the common lamiophlomis root preparation is the method for effectively controlling product quality aborning.The method of lamiophlomis rotata content is to adopt the content of general flavone (in rutin) in the spectrophotometric method detection of drugs and the content of n-butanol extract in the existing detection common lamiophlomis root preparation.Its standard is no less than 8.0% for n-butanol extract content, and general flavone (in rutin) is no less than 20.0mg.Because of the material that can be dissolved in normal butyl alcohol in the medicine more, and n-butanol extract can't form relation one to one with the content of lamiophlomis rotata in the medicine, therefore, the content that detects lamiophlomis rotata in the common lamiophlomis root preparation with the content of n-butanol extract is subjected to the interference of normal butyl alcohol soluble impurity easily.Be unfavorable for controlling aborning quality with monitor drug.
Summary of the invention
The object of the present invention is to provide the method for lamiophlomis rotata content in a kind of new detection common lamiophlomis root preparation.
Another object of the present invention just provides a kind of new standard of lamiophlomis rotata content in the common lamiophlomis root preparation that detects.
The objective of the invention is to be achieved through the following technical solutions:
Detect and to contain in the following assay one or more in the method for lamiophlomis rotata content in the common lamiophlomis root preparation:
1, determination of total flavonoids in the common lamiophlomis root preparation:
The preparation of reference substance solution: precision takes by weighing at the control substance of Rutin 0.2g of 110～130 ℃ of drying under reduced pressure to constant weight, puts in the 100ml measuring bottle, adds 60%～90% ethanol, 55～85ml, put that low-grade fever makes dissolving in the water-bath, put coldly, add 60%～80% ethanol, shake up to scale.Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and contains anhydrous rutin 0.2mg among every 1ml.
The preparation of typical curve: precision is measured reference substance solution 1.0ml, 2.0ml, 3.0ml, 4.0ml, 5.0ml, 6.0ml, puts respectively in the 25ml measuring bottle, adds water to 6ml, add 3%～10% sodium nitrite solution, 0.5～1.5ml, mixing was placed 3～10 minutes, added 6%～20% aluminum nitrate solution, 0.5～1.5ml, shake up, placed 3～10 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shake up, placed 10～20 minutes; With corresponding solution is blank.According to spectrophotometric method, measure absorbance log at 500nm wavelength place, be that ordinate, concentration are horizontal ordinate with the absorbance log, the drawing standard curve.
Determination method: the accurate title, decided common lamiophlomis root preparation (6g), places the 100ml measuring bottle, adds 60%～90% ethanol, 55～85ml, put in the water-bath low-grade fever and jolting constantly 20～40 minutes, put coldly, 60%～90% ethanol is to scale again, shake up, placed 3～5 hours, precision is measured supernatant 1ml, places the 25ml measuring bottle, add water to 6ml, add 3%～10% sodium nitrite solution, 0.5～1.5ml, mixing was placed 3～10 minutes, add 6%～20% aluminum nitrate solution, 0.5～1.5ml, shake up, placed 3～10 minutes, hydro-oxidation sodium test solution 10ml, add water to scale again, shake up, placed 10～20 minutes, according to spectrophotometric method, measure absorbance log at 500nm wavelength place, read the weight of rutin the need testing solution from typical curve, calculate, promptly.Its examination criteria is to contain general flavone with rutin (C in every 0.3g common lamiophlomis root preparation (being equivalent to crude drug 1g)
27H
30O
16) meter, must not be less than 26.0mg.
2, the assay of cyanidenon
Use high effective liquid chromatography for measuring: chromatographic condition and wavelength are selected: with octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.4% phosphoric acid solution (50: 50) is a moving phase; The detection wavelength is 350nm.Number of theoretical plate is pressed the cyanidenon peak and is calculated, and should be not less than 1500;
The preparation of reference substance solution: precision takes by weighing through the cyanidenon reference substance of phosphorus pentoxide dried overnight an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.01mg, promptly;
The preparation of need testing solution: the accurate title, decided test sample 150mg, places the 25ml measuring bottle, adds 2.5mol/L methanol hydrochloride solution 10～25ml, sonicated 20～40 minutes is put coldly, is diluted to scale with the 2.5mol/L methanol hydrochloride solution again, shake up, filter, precision is measured filtrate 10ml, put in the 50ml round-bottomed flask, heating hydrolysis is 20～40 minutes in 80～100 ℃ of water-baths, puts cold, be transferred in the 25ml measuring bottle, add methyl alcohol to scale, shake up, promptly.
Measurement result, accurate respectively reference substance solution and each 5ul of need testing solution of drawing injects high performance liquid chromatograph, measures, and promptly gets the result.Examination criteria is that the content of cyanidenon is between 0.8000mg～1.8105mg in every 0.3g common lamiophlomis root preparation (being equivalent to crude drug 1g), and promptly the content of cyanidenon is between 0.2666%～0.6035%.
Common lamiophlomis root preparation of the present invention comprises clinical acceptable forms such as capsule, tablet, granule and oral liquid, injection, film, aerosol.
Main chemical compositions in the lamiophlomis rotata is a flavone compound: Luteolin, Luteolin-7-0-glucoside, apiolin-7-0-neohesperidoside, Quercetin and Quercetin-3-0-Arabinoside.Main flavone aglycone after its hydrolysis is a Luteolin, is the main active in the lamiophlomis rotata, and content and lamiophlomis rotata content are linear under certain condition.So on basis with the colorimetric method for determining general flavone content, drug hydrolysis is measured the content of flavone aglycone Luteolin, detect the content of lamiophlomis rotata in the common lamiophlomis root preparation with this, lamiophlomis rotata content quality standard is able to further perfect, raising in the common lamiophlomis root preparation thereby make, and accomplishes the stable, controlled of common lamiophlomis root preparation quality conscientiously.
Following experimental example is used to further specify the present invention.
Experimental example 1The research of Luteolin content assaying method
(1) instrument and reagent
High performance liquid chromatograph comprises Waters 1525 pumps, 2487 binary channels UV-detector, Waters column oven, Breeze liquid chromatography workstation, Brasson ultrasonic cleaning instrument.Cyanidenon reference substance (lot number: 111520-200201, for assay usefulness, Fig. 3-5 sees in Nat'l Pharmaceutical ﹠ Biological Products Control Institute), methyl alcohol is U.S. Tedia chromatographically pure, and water is ultrapure water, and it is pure that other reagent is analysis.Drug sample to be measured is a Radix Lamiophlomidis Rotatae capsule (lot number: 0301114353 Gansu Duyiwei Biological Pharmaceutical Co., Ltd).
(2) chromatographic condition is with reference to the chromatographic condition of flavonoid of ginkgo biloba mensuration
Chromatographic column: Phenomenex Luna C
18(2), 4.6mm * 150mm, 5um; Column temperature: 35 ℃; Moving phase: methyl alcohol-0.4% phosphoric acid solution (50: 50); The detection wavelength is 350nm; Flow velocity: 0.8ml/min.
(3) selection of detection wavelength
Get the cyanidenon reference substance solution, in the interscan of 200～400nm scope, the result has absorption maximum at the 350nm place, so select 350nm as detecting wavelength.
(4) selection of hydrolysis medium: with reference to the assay method [2] of GINKGO BILOBA EXTRACT and the hydrolysising condition of licoflavone, investigate the influence of hydrolysis medium, according to the operation down of assay item to measurement result.Get the about 150mg of capsule 's content, the accurate title, decide, and puts in the 25ml measuring bottle, adds the about 20ml of methyl alcohol, sonicated 30 minutes, put coldly, be diluted to scale, shake up, filter with methyl alcohol, precision is measured subsequent filtrate 10ml, puts in the 50ml round-bottomed flask, is recycled to driedly, adds the 10ml hydrolysis medium.Hydrolysis medium is respectively methyl alcohol-25% hydrochloric acid (4: 1) and 2.5mol/L hydrochloric acid methanol, and (lot number: the content of cyanidenon 0301114353) the results are shown in Table 1 to working sample.
Table 1 hydrolysis medium is to the influence of measurement result
Hydrolysis medium content of luteolin (%) method source
Methyl alcohol-25% hydrochloric acid (4: 1) 0.281 document [2]
2.5mol/L hydrochloric acid methanol 0.390 document [3]
Therefore, select for use the 2.5mol/L hydrochloric acid methanol as hydrolysis medium.
(5) extract choice of Solvent: operate as extracting solvent, shining under the assay item with methyl alcohol, 2.5mol/L hydrochloric acid methanol respectively, (lot number: the content of cyanidenon is measured 0301114353), the results are shown in Table 2 to sample.
Table 2 extracts the influence of solvent to measurement result
Extract solvent content of luteolin (%)
Methyl alcohol 0.401
2.5mol/L hydrochloric acid methanol 0.499
Therefore, select for use the 2.5mol/L hydrochloric acid methanol as extracting solvent.
(6) selection of solvent load: add 2.5mol/L hydrochloric acid methanol 20ml respectively, 40ml sonicated 30 minutes is settled to 25ml, 50ml.Precision is measured 10ml, 20ml back hydrolysis respectively then, is settled to 25ml at last.(lot number: the content of cyanidenon is measured 0301114353), the results are shown in Table 3 to sample.
Table 3 solvent load is to the influence of measurement result
Solvent load (ml) content of luteolin (%)
20???????????????????????????????0.499
40???????????????????????????????0.498
Therefore, select for use and add 2.5mol/L hydrochloric acid methanol 20ml sonicated.
(7) selection of hydrolysis time: according to the operation down of assay item, the back hydrolysis time was respectively 0.5,1 hour, and (lot number: the content of cyanidenon 0301114353) the results are shown in Table 4 to working sample.
Table 4 hydrolysis time is to the influence of measurement result
Hydrolysis time (hour) 0.5 1
Content of luteolin (%) 0.499 0.497
Therefore, the back hydrolysis time selected for use 0.5 hour.
(8) blank test
In the ratio of prescription taste of traditional Chinese medicine, autogamy does not contain the blank preparation of lamiophlomis rotata, makes blank solution as stated above, measures in accordance with the law, and blank solution is not seen apparent chromatographic peak at the place of identical retention time with the reseda reference substance as a result, so think noiseless.
(9) linear relationship is investigated
Accurate above-mentioned reference substance solution (0.01584mg/ml) 2,4,6,8, the 10ul of drawing injects liquid chromatograph, measures peak area, is horizontal ordinate with the sample size (ug) of reference substance, is ordinate with the peak area, the drawing standard curve.The results are shown in Table 5.
Table 5 linear relationship experimental data (n=5)
Ul ug peak area
2???????????????0.03168??????????????????145187
4???????????????0.06336??????????????????296734
6???????????????0.09504??????????????????451255
8???????????????0.12672??????????????????593571
10??????????????0.15840??????????????????754208
Y＝4781815.025X-6272.7
r＝0.9999
The result shows: be good linear relationship in 0.03168～0.15840ug scope.
(10) stability test
The accurate need testing solution 5ul that draws, interval certain hour sample introduction, the peak area of mensuration cyanidenon, RSD is 0.78%, the result shows that measurement result is stable in 24 hours, the results are shown in Table 6.
Table 6 stability test data (n=5)
Sample injection time (hour) 0124 24 RSD (%)
Cyanidenon peak area 273,583 276,828 279,385 277,938 276,294 0.78
(11) precision test
The accurate need testing solution 5ul that draws injects liquid chromatograph, measures the peak area of cyanidenon, repeats sample introduction 5 times, and RSD is 0.70% as a result, sees Table 7
Table 7 precision test figure (n=5)
Sample introduction number of times 12345 RSD%
Cyanidenon peak area 273,583 272,930 276,302 276,828 277,002 0.70
(12) reappearance test
According to the operation down of assay item, (lot number: 0301114353) sample is measured for 5 parts, and trying to achieve relative standard deviation RSD is 0.87%, the results are shown in Table 8 to same lot number.
Table 8 reappearance test figure
Measure number of times content of luteolin (%) RSD%
1??????????????????0.491
2??????????????????0.495
3??????????????????0.493??????????????0.87
4??????????????????0.502
5??????????????????0.498
(13) recovery test
Reclaim with application of sample, precision takes by weighing the same lot number (lot number: 0301114353 of known content of luteolin, 0.496%) the about 75mg of sample, the accurate respectively cyanidenon reference substance solution (0.380mg/ml) that adds is a certain amount of, according to the operation down of assay item, be calculated as follows the recovery, the results are shown in Table 8 and Figure 12.
Table 9 average recovery experimental data (n=6)
The content of cyanidenon adds reference substance and is equivalent to cyanidenon and measures the cyanidenon total amount recovery in the sample sample weighting amount sample
(mg)?????????????(mg)???????????????(ml)???????????(mg)?????????????????(mg)?????????(％)
75.6??????????????0.3750??????????????0.8???????????0.304?????????????0.6720??????????97.70
71.2??????????????0.3532??????????????0.8???????????0.304?????????????0.6453??????????96.09
74.3??????????????0.3685??????????????1.0???????????0.380?????????????0.7350??????????96.45
72.8??????????????0.3611??????????????1.0???????????0.380?????????????0.7364??????????98.76
70.7??????????????0.3507??????????????1.2???????????0.456?????????????0.7933??????????97.06
70.1??????????????0.3477??????????????1.2???????????0.456?????????????0.7861??????????96.14
Average recovery rate is: 97.03%RSD is 1.08%.
Experimental example 2The assay of cyanidenon in the Radix Lamiophlomidis Rotatae capsule
According to the operation of the method for above-mentioned assay, measured the content of cyanidenon in 11 batches of Radix Lamiophlomidis Rotatae capsules, the results are shown in Table 10 (lot numbers 0301114353).
The assay of table 10 Radix Lamiophlomidis Rotatae capsule cyanidenon
Lot number content (mg/ grain) average content (mg/ grain)
0301114353????1.494????????1.485????????????1.4895
0303024151????1.056????????1.032????????????1.0440
0303024251????1.065????????1.041????????????1.0530
0303024351????1.047????????1.071????????????1.0590
0301224254????1.491????????1.530????????????1.5105
0301224251????1.509????????1.539????????????1.524
0205294121????1.827????????1.794????????????1.8105
0301114252????1.467????????1.479????????????1.4730
0301104251????1.491????????1.533????????????1.5120
0301104352????1.536????????1.548????????????1.5420
0202064321????1.554????????1.581????????????1.5675
Record every 0.3g common lamiophlomis root preparation (being equivalent to crude drug 1g) content of luteolin between 1.0440～1.8105mg, consider the loss error in the industry amplification, therefore on the basis of minimum content, float downward 20%, therefore the examination criteria that obtains is that every 0.3g common lamiophlomis root preparation (being equivalent to crude drug 1g) content of luteolin is (being that content of luteolin is between 0.2666%～0.6035%) between 0.8000mg～1.8105mg.
Experimental example 3N-butanol extract detects the defective of common lamiophlomis root preparation content
In the detection means: the n-butanol extract specificity is not strong in the existing common lamiophlomis root preparation detection method, be mainly Flavonoid substances in the chemical composition of Radix Lamiophlomidis Rotatae capsule, by to 100 batches of Radix Lamiophlomidis Rotatae capsule experimental analyses, find in its n-butanol extract, remove flavones ingredient, some uncertain composition causes and can't guarantee the accurate of testing result, and is stable.Now to wherein 10 batches analysis situation statistics is as follows: a. n-butanol extract assay method: by " Chinese pharmacopoeia version Radix Lamiophlomidis Rotatae capsule in 2000 quality standard [n-butanol extract] is measured down.B. determination of total flavonoids method in the n-butanol extract: by " Chinese pharmacopoeia version Radix Lamiophlomidis Rotatae capsule in 2000 quality standard [assay] is measured down.C. Radix Lamiophlomidis Rotatae capsule determination of total flavonoids method: by " Chinese pharmacopoeia version Radix Lamiophlomidis Rotatae capsule in 2000 quality standard [assay] is measured down.D. data statistics:
Table 11 sample title: Radix Lamiophlomidis Rotatae capsule
| Lot number | N-butanol extract | The Radix Lamiophlomidis Rotatae capsule general flavone content | General flavone content in the n-butanol extract | Uncertain composition in the n-butanol extract |
| 0307191221 | ????11.8％ | ????12.0％ | ????8.0％ | ????3.8％ |
| 0307251122 | ????11.4％ | ????14.6％ | ????9.7％ | ????1.7％ |
| 0307251323 | ????12.0％ | ????13.8％ | ????9.2％ | ????2.8％ |
| 0307271122 | ????13.0％ | ????13.7％ | ????9.1％ | ????3.9％ |
| 0307281123 | ????10.8％ | ????14.6％ | ????9.7％ | ????1.1％ |
| 0308191122 | ????12.0％ | ????13.8％ | ????9.2％ | ????2.8％ |
| 0308191222 | ????10.3％ | ????13.9％ | ????9.3％ | ????1.0％ |
| 0308201222 | ????12.3％ | ????11.2％ | ????7.5％ | ????5.0％ |
| 0308021224 | ????11.2％ | ????12.3％ | ????8.2％ | ????3.0％ |
| 0308031123 | ????12.0％ | ????13.7％ | ????9.1％ | ????2.9％ |
A. conclusion: the meltage instability of uncertain composition in the Radix Lamiophlomidis Rotatae capsule n-butanol extract, under the situation that general flavone content is more or less the same, the content of uncertain composition fluctuation is bigger in the n-butanol extract, and therefore the content with n-butanol extract detects the accuracy that lamiophlomis rotata can not be guaranteed testing result.
Test example 4The common lamiophlomis root preparation detection method relatively
The weak point that detects lamiophlomis rotata with n-butanol extract is that solvent components is indeterminate in the extract, and specificity is poor; And main chemical compositions is a flavone compound in the lamiophlomis rotata, by 30 batches of general flavone contents, all underproof Radix Lamiophlomidis Rotatae capsule of content of luteolin are measured general flavone content in its n-butanol extract, the n-butanol extract, the result shows, its n-butanol extract all meets the state-promulgated pharmacopoeia standard code, and general flavone content only accounts for 49.3% of extract total amount in the n-butanol extract.(attached Radix Lamiophlomidis Rotatae capsule n-butanol extract data analysis contrast table)
Table 12
| Lot number | General flavone content is (with rutin C 27H 30O 16Meter) standard code: every must not be less than 20.0mg | Cyanidenon (C 15H 10O 6) the content standard regulation: every must not be less than 0.8mg | N-butanol extract standard code: must not be less than 8.0% | The n-butanol extract general flavone content is (with rutin C 37H 30O 16Meter) | Uncertain composition divides content % | |
| The mg/ grain | ????％ | The mg/ grain | ||||
| ?0302112121 | ????17.5 | ????5.8 | ????0.64 | ????10.1 | ????5.0 | ????5.1 |
| ?0303102123 | ????18.4 | ????6.1 | ????0.67 | ????10.6 | ????5.3 | ????5.3 |
| ?0303032122 | ????16.5 | ????5.5 | ????0.60 | ????9.6 | ????4.8 | ????4.8 |
| ?0303110225 | ????17.6 | ????5.9 | ????0.64 | ????10.3 | ????5.1 | ????5.2 |
| ?0304022121 | ????17.0 | ????5.7 | ????0.62 | ????9.9 | ????5.0 | ????4.9 |
| ?0304232121 | ????19.2 | ????6.4 | ????0.70 | ????11.1 | ????5.6 | ????5.5 |
| ?0304132125 | ????15.7 | ????5.2 | ????0.57 | ????9.0 | ????4.5 | ????4.5 |
| ?0304152222 | ????16.9 | ????5.6 | ????0.61 | ????9.7 | ????4.9 | ????4.8 |
| ?0305022121 | ????17.8 | ????5.9 | ????0?65 | ????10.3 | ????5.1 | ????5.2 |
| ?0305202123 | ????18.0 | ????6.0 | ????0.65 | ????10.4 | ????5.2 | ????5.2 |
| ?0305212121 | ????18.4 | ????6.1 | ????0.67 | ????10.6 | ????5.3 | ????5.3 |
| ?0305202124 | ????17.8 | ????5.9 | ????0.65 | ????10.3 | ????5.1 | ????5.2 |
| ?0306212121 | ????18.1 | ????6.0 | ????0.66 | ????10.4 | ????5.2 | ????5.2 |
| ?0306232123 | ????16.5 | ????5.5 | ????0.60 | ????9.9 | ????4.8 | ????5.1 |
| ?0306252126 | ????17.4 | ????5.8 | ????0.63 | ????10.1 | ????5.0 | ????5.1 |
| ?0306282126 | ????19.4 | ????6.5 | ????0.71 | ????11.2 | ????5.6 | ????5.6 |
| ?0306292121 | ????13.8 | ????4.6 | ????0.50 | ????8.0 | ????4.0 | ????4.0 |
| ?0307022323 | ????17.7 | ????5.9 | ????0.64 | ????10.6 | ????5.1 | ????5.5 |
| ?0307052126 | ????18.0 | ????6.0 | ????0.65 | ????10.7 | ????5.2 | ????5?5 |
| ?0307092325 | ????19.5 | ????6.5 | ????0.71 | ????11.6 | ????5.7 | ????5.9 |
| ?0307132126 | ????17.8 | ????5.9 | ????0.65 | ????10.6 | ????5.1 | ????5.5 |
| ?0307202123 | ????17.1 | ????5?7 | ????0.62 | ????10.2 | ????5.0 | ????5.2 |
| ?0308122121 | ????16.5 | ????5.5 | ????0.60 | ????9.9 | ????4.8 | ????5.1 |
| ?0308152123 | ????16.8 | ????5.6 | ????0.61 | ????10.0 | ????4.9 | ????5.1 |
| ?0308202125 | ????17.3 | ????5.8 | ????0.63 | ????10.3 | ????5.0 | ????5.3 |
| ?0308232126 | ????17.2 | ????5.7 | ????0.63 | ????10.3 | ????5.0 | ????5.3 |
| ?0308292124 | ????18.7 | ????6.2 | ????0.68 | ????11.2 | ????5.4 | ????5.8 |
| ?0309012124 | ????17.8 | ????5.9 | ????0.65 | ????10.6 | ????5.1 | ????5.5 |
| ?0309032122 | ????17.0 | ????5.7 | ????0.62 | ????10.1 | ????5.0 | ????5.1 |
| ?0309082121 | ????19.7 | ????6.6 | ????0.72 | ????11.8 | ????5.7 | ????6.1 |
Experiment shows, measure the content of cyanidenon in the Radix Lamiophlomidis Rotatae capsule preparation with the HPLC method, cyanidenon can be separated (its average recovery 97.03% preferably in the sample, RSD is 1.08%), replace n-butanol extract with this method and measure, can effectively control quality aborning with monitor drug.
EmbodimentThe Radix Lamiophlomidis Rotatae capsule quality determining method
1, determination of total flavonoids in the Radix Lamiophlomidis Rotatae capsule: (Gansu Duyiwei Biological Pharmaceutical Co., Ltd's product batch number 0308201222)
The preparation of reference substance solution: precision takes by weighing at the control substance of Rutin 0.2g of 120 ℃ of drying under reduced pressure to constant weight, puts in the 100ml measuring bottle, adds 70% ethanol 70ml, puts that low-grade fever makes dissolving in the water-bath, puts coldly, adds 70% ethanol to scale, shakes up.Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and contains anhydrous rutin 0.2mg among every 1ml.
The preparation of typical curve: precision is measured reference substance solution 1.0ml, 2.0ml, 3.0ml, 4.0ml, 5.0ml, 6.0ml, puts respectively in the 25ml measuring bottle, adds water to 6ml, add 5% sodium nitrite solution 1ml, mixing was placed 6 minutes, added 10% aluminum nitrate solution 1ml, shake up, placed 6 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shake up, placed 15 minutes; With corresponding solution is blank.According to spectrophotometric method, measure absorbance log at 500nm wavelength place, be that ordinate, concentration are horizontal ordinate with the absorbance log, the drawing standard curve.
Determination method: get the content of Radix Lamiophlomidis Rotatae capsule, the accurate title, decided porphyrize, get 0.6g, the accurate title, decide, and puts in the 100ml measuring bottle, add 70% ethanol 70ml, put in the water-bath low-grade fever and jolting constantly 30 minutes, put cold, add 70% ethanol to scale, shake up, placed 4 hours, precision is measured supernatant 1ml, puts in the 25ml measuring bottle, adds water to 6ml, add 5% sodium nitrite solution 1ml, mixing was placed 6 minutes, add 10% aluminum nitrate solution 1ml, shake up, placed 6 minutes, hydro-oxidation sodium test solution 10ml, add water to scale again, shake up, placed 15 minutes, according to spectrophotometric method, measure absorbance log at 500nm wavelength place, read the weight of rutin the need testing solution from typical curve, calculate, obtain the result and contain general flavone with rutin (C in every 0.3g (being equivalent to crude drug 1.0g) Radix Lamiophlomidis Rotatae capsule
27H
30O
16) meter 33.6mg.
2, the assay of cyanidenon (Gansu Duyiwei Biological Pharmaceutical Co., Ltd produces and criticizes 0301114256)
Use high effective liquid chromatography for measuring:
Chromatographic condition and wavelength are selected: with octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.4% phosphoric acid solution (50: 50) is a moving phase; The detection wavelength is 350nm.Number of theoretical plate is pressed the cyanidenon peak and is calculated, and should be not less than 1500;
The preparation of reference substance solution: precision takes by weighing through the cyanidenon reference substance of phosphorus pentoxide dried overnight an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.01mg, promptly;
The preparation of need testing solution: get test sample content 150mg, the accurate title, decide, and puts in the 25ml measuring bottle, add 2.5mol/L methanol hydrochloride solution 20ml, sonicated 30 minutes is put cold, be diluted to scale with the 2.5mol/L methanol hydrochloride solution, shake up, filter, precision is measured filtrate 10ml, puts in the 50ml round-bottomed flask, and heating hydrolysis is 30 minutes in 90 ℃ of water-baths, put coldly, be transferred in the 25ml measuring bottle, add methyl alcohol to scale, shake up, promptly.
Measurement result: accurate respectively reference substance solution and each 5ul of need testing solution of drawing, inject high performance liquid chromatograph, measure, the content that calculates cyanidenon in every 0.3g Radix Lamiophlomidis Rotatae capsule (being equivalent to crude drug 1.0g) is 1.488mg, and promptly content of luteolin is 0.496%.
Claims (3)
1, lamiophlomis rotata content detecting method in a kind of common lamiophlomis root preparation is characterized in that containing in this method in the following assay one or more:
Determination of total flavonoids in a, the common lamiophlomis root preparation:
The preparation of reference substance solution: precision takes by weighing at the control substance of Rutin 0.2g of 110～130 ℃ of drying under reduced pressure to constant weight, places the 100ml measuring bottle, adds 60%～90% ethanol, 55～85ml, put that low-grade fever makes dissolving in the water-bath, put coldly, add 60%～80% ethanol, shake up to scale.Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and contains anhydrous rutin 0.2mg among every 1ml;
The preparation precision of typical curve is measured reference substance solution 1.0ml, 2.0ml, 3.0ml, 4.0ml, 5.0ml, 6.0ml, puts respectively in the 25ml measuring bottle, adds water to 6ml, add 3%～10% sodium nitrite solution, 0.5～1.5ml, mixing was placed 3～10 minutes, added 6%～20% aluminum nitrate solution, 0.5～1.5ml, shake up, placed 3～10 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shake up, placed 10～20 minutes; With corresponding solution is blank.According to spectrophotometric method, measure absorbance log at 500nm wavelength place, be that ordinate, concentration are horizontal ordinate with the absorbance log, the drawing standard curve;
Determination method: get common lamiophlomis root preparation, the accurate title, decide, and puts in the 100ml measuring bottle, add 60%～90% ethanol, 55～85ml, put in the water-bath low-grade fever and jolting constantly 20～40 minutes, put cold, add 60%～90% ethanol to scale, shake up, placed 3～5 hours, precision is measured supernatant 1ml, put in the 25ml measuring bottle, add water to 6ml, add 3%～10% sodium nitrite solution, 0.5～1.5ml, mixing, placed 3～10 minutes, and added 6%～20% aluminum nitrate solution, 0.5～1.5ml, shake up, placed 3～10 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shakes up, placed 10～20 minutes, according to spectrophotometric method, measure absorbance log at 500nm wavelength place, read the weight of rutin the need testing solution from typical curve, calculate, promptly.Examination criteria is to contain general flavone in rutin in every 0.3g common lamiophlomis root preparation, must not be less than 26.0mg;
The assay of b, cyanidenon:
Use high effective liquid chromatography for measuring: it is filling agent that chromatographic condition and wavelength are selected with octadecylsilane chemically bonded silica; Methyl alcohol-0.4% phosphoric acid solution is a moving phase at 50: 50; The detection wavelength is 350nm.Number of theoretical plate is pressed the cyanidenon peak and is calculated, and should be not less than 1500;
The preparation of reference substance solution: precision takes by weighing through the cyanidenon reference substance of phosphorus pentoxide dried overnight an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.01mg, promptly;
The preparation of need testing solution: the accurate title, decided common lamiophlomis root preparation 150mg, puts in the 25ml measuring bottle, adds 2.5mol/L methanol hydrochloride solution 10～25ml, sonicated 20～40 minutes is put coldly, is diluted to scale with the 2.5mol/L methanol hydrochloride solution, shake up, filter, precision is measured subsequent filtrate 10ml, put in the 50ml round-bottomed flask, heating hydrolysis is 20～40 minutes in 80～100 ℃ of water-baths, puts cold, be transferred in the 25ml measuring bottle, add methyl alcohol to scale, shake up, promptly;
Measurement result: accurate respectively reference substance solution and each 5ul of common lamiophlomis root preparation solution of drawing, inject high performance liquid chromatograph, measure, calculate, promptly.Examination criteria is that the content of cyanidenon in every 0.3g common lamiophlomis root preparation is between 0.8000mg～1.8105mg, and promptly the content of cyanidenon is between 0.2666%～0.6035%.
2, lamiophlomis rotata content detecting method as claimed in claim 1 is characterized in that containing in this method in the following assay one or more:
Determination of total flavonoids in a, the Radix Lamiophlomidis Rotatae capsule: the preparation precision of reference substance solution takes by weighing at the control substance of Rutin 0.2g of 120 ℃ of drying under reduced pressure to constant weight, puts in the 100ml measuring bottle, adds 70% ethanol 70ml, put that low-grade fever makes dissolving in the water-bath, put coldly, add 70% ethanol, shake up to scale.Precision is measured 10ml, puts in the 100ml measuring bottle, adds water to scale, shakes up, and contains anhydrous rutin 0.2mg among every 1ml;
The preparation of typical curve: precision is measured reference substance solution 1.0ml, 2.0ml, 3.0ml, 4.0ml, 5.0ml, 6.0ml, puts respectively in the 25ml measuring bottle, adds water to 6ml, add 5% sodium nitrite solution 1ml, mixing was placed 6 minutes, added 10% aluminum nitrate solution 1ml, shake up, placed 6 minutes, hydro-oxidation sodium test solution 10ml adds water to scale again, shake up, placed 15 minutes; With corresponding solution is blank.According to spectrophotometric method, measure absorbance log at 500nm wavelength place, be that ordinate, concentration are horizontal ordinate with the absorbance log, the drawing standard curve;
Determination method: get the content of Radix Lamiophlomidis Rotatae capsule, the accurate title, decided porphyrize, get 0.6g, the accurate title, decide, and puts in the 100ml measuring bottle, add 70% ethanol 70ml, put in the water-bath low-grade fever and jolting constantly 30 minutes, put cold, add 70% ethanol to scale, shake up, placed 4 hours, precision is measured supernatant 1ml, puts in the 25ml measuring bottle, adds water to 6ml, add 5% sodium nitrite solution 1ml, mixing was placed 6 minutes, add 10% aluminum nitrate solution 1ml, shake up, placed 6 minutes, hydro-oxidation sodium test solution 10ml, add water to scale again, shake up, placed 15 minutes, according to spectrophotometric method, measure absorbance log at 500nm wavelength place, read the weight of rutin the need testing solution, calculate from typical curve, obtain the result and be every 0.3g, contain general flavone in the Radix Lamiophlomidis Rotatae capsule in rutin 33.6mg;
The assay of b, cyanidenon
Use high effective liquid chromatography for measuring:
Chromatographic condition and wavelength are selected: with octadecylsilane chemically bonded silica is filling agent; Methyl alcohol-0.4% phosphoric acid solution (50: 50) is a moving phase; The detection wavelength is 350nm.Number of theoretical plate is pressed the cyanidenon peak and is calculated, and should be not less than 1500;
The preparation of reference substance solution: precision takes by weighing through the cyanidenon reference substance of phosphorus pentoxide dried overnight an amount of, adds methyl alcohol and makes the solution that every 1ml contains 0.01mg, promptly;
The preparation of need testing solution: get Radix Lamiophlomidis Rotatae capsule content 150mg, the accurate title, decide, and puts in the 25ml measuring bottle, add 2.5mol/L methanol hydrochloride solution 20ml, sonicated 30 minutes is put cold, be diluted to scale with the 2.5mol/L methanol hydrochloride solution, shake up, filter, precision is measured filtrate 10ml, puts in the 50ml round-bottomed flask, and heating hydrolysis is 30 minutes in 90 ℃ of water-baths, put coldly, be transferred in the 25ml measuring bottle, add methyl alcohol to scale, shake up, promptly;
Measurement result: accurate respectively reference substance solution and each 5ul of Radix Lamiophlomidis Rotatae capsule content solution of drawing, inject high performance liquid chromatograph, to measure, the content that calculates cyanidenon in every 0.3g Radix Lamiophlomidis Rotatae capsule is 1.488mg, promptly content of luteolin is 0.496%.
3, common lamiophlomis root preparation described in claims 1 is characterized in that formulation comprises clinical acceptable forms such as capsule, tablet, granule and oral liquid, injection, film, aerosol.
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| CN104237441B (en) * | 2014-10-09 | 2015-07-22 | 成都中医药大学 | Method for simultaneous detection of iridoid glycoside, phenylethanoid glycoside, flavone and dicaffeoyl ingredients in lamiophlomis rotata |
| CN105343162A (en) * | 2015-12-01 | 2016-02-24 | 徐斌 | Preparation method and detection method of lamiophlomis rotata kudo oral slow-release adhesive film |
| CN106093249A (en) * | 2016-08-25 | 2016-11-09 | 南京林业大学 | A kind of method determining Carpinus betulus leaf collection period |
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