CN101576545B - Method for determining content of compound Caoshanhu lozenge - Google Patents
Method for determining content of compound Caoshanhu lozenge Download PDFInfo
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- CN101576545B CN101576545B CN200910114907.XA CN200910114907A CN101576545B CN 101576545 B CN101576545 B CN 101576545B CN 200910114907 A CN200910114907 A CN 200910114907A CN 101576545 B CN101576545 B CN 101576545B
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Abstract
The invention relates to a method for determining the content of compound Caoshanhu lozenge prepared from glabrous sarcandra herb extractum, menthol and dementholised peppermint oil. The thin-layer chromatography is used for distinguishing the menthol and the glabrous sarcandra herb extractum in the lozenge. The method is characterized in that the high performance liquid chromatography is used forsimultaneously determining the content of two marker components, including isofraxidin and rosmarinic acid in the lozenge under the chromatographic condition of isocratic elution; or the high perform ance liquid chromatography is used for simultaneously testing the content of four marker components, including protocatechuic acid, caffeic acid, the isofraxidin and the rosmarinic acid in the lozengeunder the chromatographic condition of gradient elution.
Description
Technical field
The present invention relates to a kind of content assaying method of compound Chinese medicinal preparation, particularly relate to the content assaying method of Chinese medicine compound prescription Caoshan Hu Hanpian.
Background technology
The compound caoshanhu tablets that the Pharmacopoeia of the People's Republic of China (one of version in 2005) records is with Glabrous Sarcandra Herb medicinal extract, menthol, peppermint oil dementholized is the compound Chinese medicinal preparation that raw material is prepared from, [method for making] above three tastes, Glabrous Sarcandra Herb is got by Glabrous Sarcandra Herb medicinal extract system, boiling secondary, 2 hours for the first time, 1.5 hours for the second time, collecting decoction, filter, it is 1.15 (80 ℃) that filtrate is concentrated into relative density, add ethanol to reaching 65% containing alcohol amount, leave standstill 24 hours, filter, decompression filtrate recycling ethanol, and to be condensed into relative density be 1.24～1.26 clear cream, obtain.Get Glabrous Sarcandra Herb medicinal extract, add right amount of auxiliary materials, granulation, dry; Menthol is mixed with peppermint oil dementholized and make to dissolve, mix with above-mentioned particle, compacting is in blocks, or film coating, to obtain final product small pieces 0.44g, large stretch of 1.0g.There is effect of wind and heat dispersing, swelling and pain relieving, clearing throat, clinical in larynx numbness, acute laryngopharyngitis etc. due to affection of exogenous wind-heat.
The primary raw material of compound caoshanhu tablets is Glabrous Sarcandra Herb medicinal extract, is Chinese medicinal material of sarcandra glaber concentrated forming after water extract-alcohol precipitation, has wherein comprised the chemical composition that a large amount of polarity is stronger.As Huang Mingju, Li Yanlan etc. just separate and have obtained Rosmarinic acid, methyl rosmarinate, caffeic acid, danshensu first methyl esters, 3 from Glabrous Sarcandra Herb water soluble part, 4-dihydroxy-benzoic acid, isofraxidin, 7 compounds of dibutyl phthalate, and above-claimed cpd mostly has to compound caoshanhu tablets and cures mainly the pharmacologically active that function is relevant.As PCA has antibacterial action, when in vitro test, Pseudomonas aeruginosa, Escherichia coli, typhoid bacillus, shigella dysenteriae, Bacillus alcaligenes and hay bacillus and staphylococcus aureus are all had to bacteriostasis in various degree; Caffeic acid has antibacterial, antiviral, central excitation, removing toxic substances, the purposes such as blood coagulation; The effects such as Rosmarinic acid has anti-inflammation according to the literature, antitumor, inhibition blood platelet reduces, adjusting immunity of organism, very close with the pharmacologically active of Chloranthus glaber herb.
But the method for quality control that compound caoshanhu tablets adopts in 2005 editions standards of pharmacopoeia has been differentiated menthol and Glabrous Sarcandra Herb medicinal extract wherein except [discriminating] adopts thin-layered chromatography, in [assay] item, only measure the wherein content of isofraxidin, although according to the literature isofraxidin have certain antitumor action and its content in this lozenge higher, do not report that it has antibacterial, anti-inflammatory etc. and cures mainly relevant pharmacologically active to compound caoshanhu tablets function.So, only use isofraxidin as surveying index containing in this lozenge method of quality control, it contains surveys the quality good or not that result can not well reflect this lozenge.The correlativity curing mainly take the pharmacologically active of index components and the function of preparation and the content of index components in preparation are as according to selecting containing surveying index, and the mode that adopts many indexs simultaneously to control, will further strengthen rationality and the reliability of method of quality control.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, the method of quality control of the compound caoshanhu tablets that a kind of controllability is stronger is provided, be primarily characterized in that the content assaying method that has adopted multi-target ingredient simultaneously to control, and selecting, containing surveying when index take the pharmacologically active of index components and the correlativity being cured mainly by the function of controlling agent and the content of index components in preparation as foundation, to have strengthened rationality and the reliability of whole method of quality control.
The present invention is achieved by the following technical solutions:
The method of quality control of compound caoshanhu tablets of the present invention is the improvement to the method for quality control that in 2005 editions standards of pharmacopoeia, compound caoshanhu tablets adopts, comprise with thin-layered chromatography differentiate menthol wherein and Glabrous Sarcandra Herb medicinal extract, with the wherein content of index components of high effective liquid chromatography for measuring.Wherein differentiate that by thin-layered chromatography step and the former method of menthol wherein and Glabrous Sarcandra Herb medicinal extract are basic identical, only developping agent used changes sherwood oil (30～60 ℃)-ethyl acetate (9: 1) into when differentiating menthol; It is characterized in that wherein under the chromatographic condition of isocratic elution, measuring the content of wherein isofraxidin and 2 index components of Rosmarinic acid when the content of index components simultaneously or under the chromatographic condition of gradient elution, measuring the wherein content of at least 2 index components simultaneously with high effective liquid chromatography for measuring, described index components is selected from protocatechuic acid, caffeic acid, isofraxidin, Rosmarinic acid.
Comprehensive current document analysis, separates the compound obtaining from Glabrous Sarcandra Herb water soluble part, and pharmacologically active and the function of compound caoshanhu tablets cure mainly more relevant Rosmarinic acid, caffeic acid, the protocatechuic acid of mainly containing, wherein the highest with the content of Rosmarinic acid again; Although the correlativity that the pharmacologically active of isofraxidin and the function of compound caoshanhu tablets cure mainly is lower, because its content in Glabrous Sarcandra Herb is higher, so be used as its characteristic component for the various quality controls containing 'Zhongjiefeng ' always.Therefore, in order to improve the quality control standard of compound caoshanhu tablets, method of quality control of the present invention can adopt and measure 2 content compared with the method for high target composition (as isofraxidin and Rosmarinic acid) simultaneously in [assay] item, also can adopt the wherein method of at least 2 any index of correlation compositions (being selected from isofraxidin, Rosmarinic acid, caffeic acid, protocatechuic acid) of simultaneously measuring.
According to the present invention, can under the chromatographic condition of isocratic elution, measure compound caoshanhu tablets by high performance liquid chromatography simultaneously in isofraxidin and 2 content of Rosmarinic acid compared with the content of high target composition, its concrete steps are as follows:
(1) chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take proportioning as 30～20: 70～80 acetonitrile-0.1% phosphoric acid solution is mobile phase; Detection wavelength is 270--344nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak.
(2) isofraxidin reference substance is got in the preparation of reference substance solution, Rosmarinic acid reference substance is appropriate, accurately weighed, adds methyl alcohol or ethanol or acetonitrile and makes every 1ml respectively containing the solution of 5～20 μ g, to obtain final product.
(3) 10～40 of this product are got in the preparation of need testing solution, accurately weighed, porphyrize, get approximately 1～4g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol or ethanol or acetonitrile 25～100ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30～60 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol or ethanol or acetonitrile, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and each 5～20 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
The content assaying method of isofraxidin and 2 index components of Rosmarinic acid in simultaneously measuring compound caoshanhu tablets according to the above-mentioned chromatographic condition at isocratic elution, its preferred steps is as follows:
(1) chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile-0.1% phosphoric acid solution (20: 80) as mobile phase; Detection wavelength is 342nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak.
(2) isofraxidin reference substance is got in the preparation of reference substance solution, Rosmarinic acid reference substance is appropriate, accurately weighed, adds methyl alcohol and makes every 1ml respectively containing the solution of 8 μ g, 40 μ g, to obtain final product.
(3) 10 of this product are got in the preparation of need testing solution, accurately weighed, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 40.0 μ g, and sheet must not be less than 100.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 280.0 μ g, and sheet must not be less than 650.0 μ g.
Technical scheme of the present invention can also be, measures the content of compound caoshanhu tablets protocatechuic acid, caffeic acid, isofraxidin, 4 index components of Rosmarinic acid by high performance liquid chromatography under the chromatographic condition of gradient elution simultaneously, and its concrete steps are as follows:
(1) chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Mobile phase A is acetonitrile, and Mobile phase B is 0.1% phosphoric acid (containing 2.5% acetonitrile); Flow velocity is per minute 1.0ml, linear gradient elution; Column temperature is 25～38 ℃; Detection wavelength is 270～300nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak, and the degree of separation at protocatechuic acid peak, caffeic acid, isofraxidin peak and Rosmarinic acid peak is all not less than 2.0.Eluent gradient variation sees the following form:
(2) it is appropriate that protocatechuic acid, caffeic acid, isofraxidin, Rosmarinic acid reference substance are got respectively in the preparation of reference substance solution, accurately weighed, add first or ethanol or acetonitrile alcohol and make the mixed solution of every 1ml containing protocatechuic acid 1～3 μ g, caffeic acid 15～20 μ g, isofraxidin 5～10 μ g and Rosmarinic acid 30～40 μ g, to obtain final product.
(3) 10～40 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get approximately 1～4g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol or ethanol or acetonitrile 25～100ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30～60 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol or ethanol or acetonitrile, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and each 5～20 μ l of need testing solution of drawing respectively, inject high performance liquid chromatograph, measure, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with protocatechuic acid (C
7h
6o
4) meter, small pieces must not be less than 5.0 μ g, and sheet must not be less than 6.0 μ g; With caffeic acid (C
8h
8o
4) meter, small pieces must not be less than 30 μ g, and sheet must not be less than 60.0 μ g; With isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
According to above-mentioned under the chromatographic condition of gradient elution, measure compound caoshanhu tablets simultaneously in the content assaying method of isofraxidin, Rosmarinic acid, caffeic acid, 4 index components of protocatechuic acid, its preferred steps is as follows:
(1) chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Mobile phase A is acetonitrile, and Mobile phase B is 0.1% phosphoric acid containing 2.5% acetonitrile; Flow velocity is per minute 1.0ml, linear gradient elution; Column temperature is 38 ℃; Detection wavelength is 294nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak, and the degree of separation at protocatechuic acid peak, caffeic acid, isofraxidin peak and Rosmarinic acid peak is all not less than 2.0.Eluent gradient variation sees the following form:
(2) it is appropriate that protocatechuic acid, caffeic acid, isofraxidin, Rosmarinic acid reference substance are got respectively in the preparation of reference substance solution, accurately weighed, add methyl alcohol and make the mixed solution of every 1ml containing protocatechuic acid 1.5 μ g, caffeic acid 18 μ g, isofraxidin 7.5 μ g and Rosmarinic acid 40 μ g, to obtain final product.
(3) 10 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, inject high performance liquid chromatograph, measure, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with protocatechuic acid (C
7h
6o
4) meter, small pieces must not be less than 5.0 μ g, and sheet must not be less than 6.0 μ g; With caffeic acid (C
8h
8o
4) meter, small pieces must not be less than 30.0 μ g, and sheet must not be less than 60.0 μ g; With isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
According to the present invention, in can measuring compound caoshanhu tablets under the chromatographic condition at gradient elution by high performance liquid chromatography on the basis of isofraxidin, Rosmarinic acid, caffeic acid, 4 index components content of protocatechuic acid simultaneously, according to specific needs, can select wherein any 2 or 3 index components as the content control composition in actual mass control method, and high-efficient liquid phase chromatogram condition used is constant.
Compared with the prior art compound caoshanhu tablets method of quality control of the present invention has the following advantages:
(1) in assay of the present invention, select to contain survey index take the pharmacologically active of index components and the correlativity being cured mainly by the function of controlling agent and the content of index components in preparation as foundation, strengthened rationality and the reliability of compound caoshanhu tablets method of quality control.
(2) in assay of the present invention, adopt multi-target ingredient method for measuring simultaneously, further strengthened the controllability of compound caoshanhu tablets method of quality control.
(3) in assay of the present invention, adopted the need testing solution preparation method of the ultrasonic direct extraction of methyl alcohol, changed the mode that need testing solution preparation in the past adopts methenyl choloride repeatedly to extract, method is easy, favorable reproducibility, environmental pollution are little.
Accompanying drawing explanation
The high-efficient liquid phase chromatogram of assay in Fig. 1 embodiment 1.
The high-efficient liquid phase chromatogram of assay in Fig. 2 embodiment 2.
Embodiment
Below in conjunction with specific embodiment, the invention will be further described, but the present invention is not limited in cited embodiment.
Embodiment 1
A content assaying method for compound caoshanhu tablets, comprises and differentiates menthol and Glabrous Sarcandra Herb medicinal extract wherein, under the chromatographic condition of isocratic elution, measures the wherein content of isofraxidin and 2 index components of Rosmarinic acid by high performance liquid chromatography simultaneously by thin-layered chromatography.Concrete steps comprise:
[assay]
(1) chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile-0.1% phosphoric acid solution (20: 80) as mobile phase; Detection wavelength is 342nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak.
(2) isofraxidin reference substance is got in the preparation of reference substance solution, Rosmarinic acid reference substance is appropriate, accurately weighed, adds methyl alcohol and makes every 1ml respectively containing the solution of 8 μ g, 40 μ g, to obtain final product.
(3) 10 of this product are got in the preparation of need testing solution, accurately weighed, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 40.0 μ g, and sheet must not be less than 100.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 280.0 μ g, and sheet must not be less than 650.0 μ g.
Embodiment 2
A content assaying method for compound caoshanhu tablets, comprises and differentiates menthol and Glabrous Sarcandra Herb medicinal extract wherein, under the chromatographic condition of isocratic elution, measures the wherein content of isofraxidin and 2 index components of Rosmarinic acid by high performance liquid chromatography simultaneously by thin-layered chromatography.Concrete steps comprise:
[assay]
(1) chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take acetonitrile-0.1% phosphoric acid solution (30: 70) as mobile phase; Detection wavelength is 270nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak.
(2) isofraxidin reference substance is got in the preparation of reference substance solution, Rosmarinic acid reference substance is appropriate, accurately weighed, adds methyl alcohol and makes every 1ml respectively containing the solution of 10 μ g, to obtain final product.
(3) 40 of this product are got in the preparation of need testing solution, accurately weighed, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision adds ethanol 100ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30 minutes, lets cool, weighed weight again, supplies the weight of less loss with ethanol, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
Embodiment 3
A content assaying method for compound caoshanhu tablets is measured the content of its protocatechuic acid, caffeic acid, isofraxidin, 4 index components of Rosmarinic acid under the chromatographic condition of gradient elution simultaneously by high performance liquid chromatography, concrete steps comprise:
[assay]
(1) chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Mobile phase A is acetonitrile, and Mobile phase B is 0.1% phosphoric acid (containing 2.5% acetonitrile); Flow velocity is per minute 1.0ml, linear gradient elution; Column temperature is 38 ℃; Detection wavelength is 294nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak, and the degree of separation at protocatechuic acid peak, caffeic acid, isofraxidin peak and Rosmarinic acid peak is all not less than 2.0.Eluent gradient changes eluent gradient and is changed to 0 to 35min, mobile phase A: 0 to 16%; Mobile phase B: 100 to 84%; 35 to 60min, mobile phase A: 16 to 70%; Mobile phase B: 84 to 30%;
(2) it is appropriate that protocatechuic acid, caffeic acid, isofraxidin, Rosmarinic acid reference substance are got respectively in the preparation of reference substance solution, accurately weighed, add methyl alcohol and make the mixed solution of every 1ml containing protocatechuic acid 1.5 μ g, caffeic acid 18 μ g, isofraxidin 7.5 μ g and Rosmarinic acid 40 μ g, to obtain final product.
(3) 10 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, inject high performance liquid chromatograph, measure, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with protocatechuic acid (C
7h
6o
4) meter, small pieces must not be less than 5.0 μ g, and sheet must not be less than 6.0 μ g; With caffeic acid (C
8h
8o
4) meter, small pieces must not be less than 30.0 μ g, and sheet must not be less than 60.0 μ g; With isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
Embodiment 4
A content assaying method for compound caoshanhu tablets is measured the content of its protocatechuic acid, caffeic acid, isofraxidin, 4 index components of Rosmarinic acid under the chromatographic condition of gradient elution simultaneously by high performance liquid chromatography, concrete steps comprise:
[assay]
(1) chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Mobile phase A is acetonitrile, and Mobile phase B is 0.1% phosphoric acid (containing 2.5% acetonitrile); Flow velocity is per minute 1.0ml, linear gradient elution; Column temperature is 38 ℃; Detection wavelength is 294nm.Number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak, and the degree of separation at protocatechuic acid peak, caffeic acid, isofraxidin peak and Rosmarinic acid peak is all not less than 2.0.Eluent gradient changes eluent gradient and is changed to 0 to 35min, mobile phase A: 0 to 16%; Mobile phase B: 100 to 84%; 35 to 60min, mobile phase A: 16 to 70%; Mobile phase B: 84 to 30%;
(2) it is appropriate that protocatechuic acid, caffeic acid, isofraxidin, Rosmarinic acid reference substance are got respectively in the preparation of reference substance solution, accurately weighed, add methyl alcohol or ethanol or acetonitrile and make the mixed solution of every 1ml containing protocatechuic acid 1.5 μ g, caffeic acid 18 μ g, isofraxidin 7.5 μ g and Rosmarinic acid 40 μ g, to obtain final product.
(3) 40 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision add methyl alcohol or or ethanol or acetonitrile 100ml, close plug, weighed weight, ultrasonic processing (power 250W, frequency 25kHz) 30 minutes, lets cool, weighed weight again, with methyl alcohol or or ethanol or acetonitrile supply the weight of less loss, shake up, filter, get subsequent filtrate, to obtain final product.
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, inject high performance liquid chromatograph, measure, and to obtain final product.
Every of this product contains Glabrous Sarcandra Herb with protocatechuic acid (C
7h
6o
4) meter, small pieces must not be less than 5.0 μ g, and sheet must not be less than 6.0 μ g; With caffeic acid (C
8h
8o
4) meter, small pieces must not be less than 30.0 μ g, and sheet must not be less than 60.0 μ g; With isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
Claims (4)
1. the content assaying method of the compound caoshanhu tablets being formed by Glabrous Sarcandra Herb medicinal extract, menthol and peppermint oil dementholized, it is characterized in that measuring under the chromatographic condition of isocratic elution by high performance liquid chromatography the wherein content of isofraxidin and 2 index components of Rosmarinic acid, the concrete steps of assay are as follows simultaneously:
(1) chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take proportioning as 30～20: 70～80 acetonitrile-0.1% phosphoric acid solution is mobile phase; Detection wavelength is 270～344nm, and number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak;
(2) isofraxidin reference substance is got in the preparation of reference substance solution, Rosmarinic acid reference substance is appropriate, accurately weighed, adds methyl alcohol or ethanol or acetonitrile and makes every 1ml respectively containing the solution of 5～20 μ g, to obtain final product;
(3) 10～40 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get 1～4g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol or ethanol or acetonitrile 25～100ml, close plug, weighed weight, use power 250W, the ultrasonic processing of frequency 25kHz 30～60 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol or ethanol or acetonitrile, shake up, filter, get subsequent filtrate, to obtain final product;
(4) determination method accurate reference substance solution and each 5～20 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures, and to obtain final product;
Every of this product contains Glabrous Sarcandra Herb with isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
2. the content assaying method of the compound caoshanhu tablets being formed by Glabrous Sarcandra Herb medicinal extract, menthol and peppermint oil dementholized, it is characterized in that measuring under the chromatographic condition of isocratic elution by high performance liquid chromatography the wherein content of isofraxidin and 2 index components of Rosmarinic acid, the concrete steps of assay are as follows simultaneously:
(1) chromatographic condition and system suitability are take octadecylsilane chemically bonded silica as filling agent; Take proportioning as acetonitrile-0.1% phosphoric acid solution of 20: 80 is as mobile phase; Detection wavelength is 342nm, and number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak;
(2) isofraxidin reference substance is got in the preparation of reference substance solution, Rosmarinic acid reference substance is appropriate, accurately weighed, adds methyl alcohol and makes every 1ml respectively containing the solution of 8 μ g, 40 μ g, to obtain final product;
(3) 10 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get 1g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, use power 250W, the ultrasonic processing of frequency 25kHz 30 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product;
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, injection liquid chromatography, measures, and to obtain final product;
Every of this product contains Glabrous Sarcandra Herb with isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 40.0 μ g, and sheet must not be less than 100.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 280.0 μ g, and sheet must not be less than 650.0 μ g.
3. the content assaying method of the compound caoshanhu tablets being formed by Glabrous Sarcandra Herb medicinal extract, menthol and peppermint oil dementholized, the content that it is characterized in that simultaneously measuring under the chromatographic condition of gradient elution by high performance liquid chromatography its protocatechuic acid, caffeic acid, isofraxidin, 4 index components of Rosmarinic acid, the concrete steps of assay are as follows:
(1) chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Mobile phase A is acetonitrile, and Mobile phase B is 0.1% phosphoric acid containing 2.5% acetonitrile; Flow velocity is per minute 1.0ml, linear gradient elution; Column temperature is 25～38 ℃; Detection wavelength is 270～300nm, number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak, the degree of separation at former several boheic acids peak, caffeic acid, isofraxidin peak and Rosmarinic acid peak is all not less than 2.0, and eluent gradient is changed to 0 to 35min, mobile phase A: 0 to 16%; Mobile phase B: 100 to 84%; 35 to 60min, mobile phase A: 16 to 70%; Mobile phase B: 84 to 30%;
(2) it is appropriate that protocatechuic acid, caffeic acid, isofraxidin, Rosmarinic acid reference substance are got respectively in the preparation of reference substance solution, accurately weighed, add methyl alcohol or ethanol or acetonitrile and make the mixed solution of every 1ml containing protocatechuic acid 1～3 μ g, caffeic acid 15～20 μ g, isofraxidin 5～10 μ g and Rosmarinic acid 30～40 μ g, to obtain final product;
(3) 10～40 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get 1～4g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol or ethanol or acetonitrile 25～100ml, close plug, weighed weight, use power 250W, the ultrasonic processing of frequency 25kHz 30～60 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol or ethanol or acetonitrile, shake up, filter, get subsequent filtrate, to obtain final product;
(4) determination method accurate reference substance solution and each 5～20 μ l of need testing solution of drawing respectively, inject high performance liquid chromatograph, measure, and to obtain final product;
Every of this product contains Glabrous Sarcandra Herb with protocatechuic acid (C
7h
6o
4) meter, small pieces must not be less than 5.0 μ g, and sheet must not be less than 6.0 μ g; With caffeic acid (C
8h
8o
4) meter, small pieces must not be less than 30 μ g, and sheet must not be less than 60.0 μ g; With isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
4. content assaying method according to claim 3, it is characterized in that: under the chromatographic condition of gradient elution, measure the content of its protocatechuic acid, caffeic acid, isofraxidin, 4 index components of Rosmarinic acid by high performance liquid chromatography, the concrete steps of assay are as follows simultaneously:
(1) chromatographic condition and system suitability octadecylsilane chemically bonded silica are filling agent; Mobile phase A is acetonitrile, and Mobile phase B is 0.1% phosphoric acid containing 2.5% acetonitrile; Flow velocity is per minute 1.0ml, linear gradient elution; Column temperature is 38 ℃; Detection wavelength is 294nm, and number of theoretical plate calculates and should be not less than 1200 by isofraxidin peak, and the degree of separation at protocatechuic acid peak, caffeic acid, isofraxidin peak and Rosmarinic acid peak is all not less than 2.0, and eluent gradient is changed to 0 to 35min, mobile phase A: 0 to 16%; Mobile phase B: 100 to 84%; 35 to 60min, mobile phase A: 16 to 70%; Mobile phase B: 84 to 30%;
(2) it is appropriate that protocatechuic acid, caffeic acid, isofraxidin, Rosmarinic acid reference substance are got respectively in the preparation of reference substance solution, accurately weighed, add methyl alcohol and make the mixed solution of every 1ml containing protocatechuic acid 1.5 μ g, caffeic acid 18 μ g, isofraxidin 7.5 μ g and Rosmarinic acid 40 μ g, to obtain final product;
(3) 10 of this product are got in the preparation of need testing solution, remove film-coating, accurately weighed, porphyrize, get about 1g, accurately weighed, put in tool plug conical flask, precision adds methyl alcohol 25ml, close plug, weighed weight, use power 250W, the ultrasonic processing of frequency 25kHz 30 minutes, lets cool, weighed weight again, supplies the weight of less loss with methyl alcohol, shake up, filter, get subsequent filtrate, to obtain final product;
(4) determination method accurate reference substance solution and the each 10 μ l of need testing solution of drawing respectively, inject high performance liquid chromatograph, measure, and to obtain final product;
Every of this product contains Glabrous Sarcandra Herb with protocatechuic acid (C
7h
6o
4) meter, small pieces must not be less than 5.0 μ g, and sheet must not be less than 6.0 μ g; In caffeic acid (C8H8O4), small pieces must not be less than 30.0 μ g, and sheet must not be less than 60.0 μ g; With isofraxidin (C
11h
10o
5) meter, small pieces must not be less than 20.0 μ g, and sheet must not be less than 50.0 μ g; With Rosmarinic acid (C
18h
16o
8) meter, small pieces must not be less than 100.0 μ g, and sheet must not be less than 250.0 μ g.
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| CN104897789B (en) * | 2015-03-17 | 2018-07-13 | 广西昌弘制药有限公司 | A kind of method of quality control of the preparation of relieving sore throat and acesodyne |
| CN107007655A (en) * | 2017-04-14 | 2017-08-04 | 佛山市海科知识产权交易有限公司 | A kind of compound Caoshanhu thoat-soothing drops and preparation method thereof |
| CN107137647A (en) * | 2017-06-25 | 2017-09-08 | 长沙善道新材料科技有限公司 | A kind of compound Caoshanhu thoat-soothing drops and preparation method thereof |
| CN111721880B (en) * | 2020-07-02 | 2023-01-10 | 江中药业股份有限公司 | Method for establishing fingerprint of sarcandra glabra by using double-column tandem HPLC-MS |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1985862A (en) * | 2006-12-15 | 2007-06-27 | 中国科学院长春应用化学研究所 | Quality analysis and identification of herbal medicine material glabrous sarcandra herb and Chinese medicine therewith |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1985862A (en) * | 2006-12-15 | 2007-06-27 | 中国科学院长春应用化学研究所 | Quality analysis and identification of herbal medicine material glabrous sarcandra herb and Chinese medicine therewith |
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| 黄增琼、蒋伟哲等.高效液相色谱法测定肿节风含片中异嗪皮啶含量.《药物鉴定》.2007,第16卷(第5期),19-20. |
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