CN106198823A - A kind of assay method of SHENSHUAINING sheet effective ingredient - Google Patents
A kind of assay method of SHENSHUAINING sheet effective ingredient Download PDFInfo
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- CN106198823A CN106198823A CN201610712681.3A CN201610712681A CN106198823A CN 106198823 A CN106198823 A CN 106198823A CN 201610712681 A CN201610712681 A CN 201610712681A CN 106198823 A CN106198823 A CN 106198823A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/86—Signal analysis
- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
- G01N30/8686—Fingerprinting, e.g. without prior knowledge of the sample components
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- 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
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Abstract
nullThe present invention relates to the assay method of a kind of SHENSHUAINING sheet effective ingredient，Described method comprises the steps: step 1: reference substance solution preparation takes danshensu sodium、Salvianolic acid B、Hesperidin、Berberine hydrochloride、Aloe-emodin、Rheum emodin、Chrysophanic acid、Chrysophanol、Physcione reference substance is appropriate，Accurately weighed，Add methanol and be respectively prepared solution，Step 2: the preparation of control medicinal material solution takes Radix Pseudostellariae、Rhizoma Coptidis、The Rhizoma Pinelliae、Pericarpium Citri Reticulatae、Poria、Radix Et Rhizoma Rhei、Radix Salviae Miltiorrhizae、Radix Achyranthis Bidentatae、Flos Carthami、Licorice medicinal materials powder，Accurately weighed，Add methanol and make solution，Step 3: need testing solution preparation takes sample powder，Accurately weighed，Add methanol and make solution，Step 4: algoscopy takes each 5 15 μ L of above-mentioned solution respectively，It is injected separately into chromatograph of liquid detection，Record chromatogram，Gained chromatogram is compared with standard finger-print.
Description
Technical field
The present invention relates to a kind of traditional Chinese medicine test method, particularly to the assay method of a kind of SHENSHUAINING sheet effective ingredient.
Background technology
SHENSHUAINING sheet is a kind of Chinese patent medicine listed, and formula is: Radix Pseudostellariae, Rhizoma Coptidis, Rhizoma Pinelliae (processed), Pericarpium Citri Reticulatae, Poria,
Radix Et Rhizoma Rhei, Radix Salviae Miltiorrhizae, Radix Achyranthis Bidentatae, Flos Carthami, Radix Glycyrrhizae.
SHENSHUAINING sheet has replenishing QI to invigorate the spleen, blood circulation promoting and blood stasis dispelling, and purging FU-organs lets out turbid effect.For spleen weakness, the turbid retardance of the stasis of blood, lifting
Lumbago caused by imbalance is tired, shallow complexion, nausea and vomiting, inappetence, dysuria, and stool viscous and many reasons draw
The chronic renal insufficiency risen is shown in above-mentioned disease person.
The detection method of existing SHENSHUAINING sheet specifically includes that the discriminating of Radix Et Rhizoma Rhei, the discriminating of Rhizoma Coptidis, the discriminating of Radix Salviae Miltiorrhizae, and
The mensuration of content of berberine hydrochloride, and content of Danshensu mensuration, wherein berberine hydrochloride content method uses spectrophotometric
Method, measures trap at the wavelength of 345nm, calculates content by the trap of berberine hydrochloride.Wherein content of Danshensu measures
Use high performance liquid chromatography, select the effective ingredient danshensu of Radix Salviae Miltiorrhizae in prescription as content's index, to use HPLC method to carry out
Measure, chromatographic condition flowing phase: phosphate buffer (0.5%Nail2P04-0.03%H3P04): methanol=95:5；Detection ripple
Long 220nm；Flow velocity: lml/ml；Chromatographic column: Purospher STAR RP mono-18e, 5um (1504.6mn)；Post
But owing to SHENSHUAINING sheet chemical composition kind is more and physicochemical property differs greatly, only minority composition in preparation is entered
The quality of the method evaluation preparation that row is quantitative is the most unilateral；It is difficult to its chemical feature of complete characterization.
Chinese medicine fingerprint from the complete chemical composition embodying Chinese medicine of analytic angle, thus can realize matter to Chinese medicine
Amount controls.Hence set up SHENSHUAINING sheet finger printing, the total quality of preparation can be reflected the most intuitively, all sidedly.Pin of the present invention
To Chinese medicine preparation SHENSHUAINING sheet, establish the evaluation methodology of finger printing, study and realize the overall matter to preparation with system fingerprint
Amount is evaluated.
Summary of the invention:
The invention discloses the detection method of a kind of SHENSHUAINING sheet, described method comprises the steps:
Step 1: prepared by reference substance solution
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, adds methanol and is respectively prepared every 0.8-1.2ml containing danshensu sodium, berberine hydrochloride
45-55 μ g, containing salvianolic acid B, Hesperidin 80-120 μ g, containing aloe-emodin, rheum emodin, chrysophanol, physcione 8-12 μ g
Solution, to obtain final product.
Step 2: prepared by control medicinal material solution
Take Radix Pseudostellariae (0.4-0.6g), Rhizoma Coptidis (0.18-0.22g), the Rhizoma Pinelliae (0.4-0.6g), Pericarpium Citri Reticulatae (0.18-respectively
0.22g), Poria (0.3-0.5g), Radix Et Rhizoma Rhei (0.7-0.9g), Radix Salviae Miltiorrhizae (0.8-1.2g), Radix Achyranthis Bidentatae (0.3-0.5g), Flos Carthami (0.18-
0.22g), Radix Glycyrrhizae (0.18-0.22g) medicinal powder, accurately weighed, put in tool plug conical flask, accurate add 70-90% methanol
20-30ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, supply with 80% methanol and subtract
The weight lost, shakes up, and filters, takes subsequent filtrate, to obtain final product.
Step 3: prepared by need testing solution
Take sample powder 0.8-1.2g, accurately weighed, put in tool plug conical flask, accurate addition 45-55% methanol 45-
55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, supply less loss with 50% methanol
Weight, shake up, filter, take subsequent filtrate, to obtain final product.
Step 4: algoscopy
Take reference substance solution and need testing solution each 5-15 μ L respectively, be injected separately into chromatograph of liquid detection, record chromatograph
Figure, compares gained chromatogram with standard finger-print.
Preferably, the detection method of the present invention, comprise the steps:
Step 1: prepared by reference substance solution
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, add methanol be respectively prepared every 1ml containing danshensu sodium, berberine hydrochloride 50 μ g, contain
Salvianolic acid B, Hesperidin 100 μ g, containing aloe-emodin, rheum emodin, chrysophanol, the solution of physcione 10 μ g, to obtain final product.
Step 2: prepared by control medicinal material solution
Take Radix Pseudostellariae (0.5g), Rhizoma Coptidis (0.2g), the Rhizoma Pinelliae (0.5g), Pericarpium Citri Reticulatae (0.2g), Poria (0.4g), Radix Et Rhizoma Rhei respectively
(0.8g), Radix Salviae Miltiorrhizae (1.0g), Radix Achyranthis Bidentatae (0.4g), Flos Carthami (0.2g), the medicinal powder such as Radix Glycyrrhizae (0.2g), accurately weighed, put tool plug
In conical flask, accurate addition 80% methanol 25ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 points
Clock, takes out, lets cool, more weighed weight, supplies the weight of less loss with 80% methanol, shakes up, and filters, takes subsequent filtrate, to obtain final product.
Step 3: prepared by need testing solution
Take the sample powder under assay item, take about 1.0g, accurately weighed, put in tool plug conical flask, accurate addition
50% methanol 50ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 minutes, take out, let cool, then claim
Determine weight, supply the weight of less loss with 50% methanol, shake up, filter, take subsequent filtrate, to obtain final product.
Step 4: algoscopy
Take reference substance solution and need testing solution each 1-6 μ L respectively, be injected separately into chromatograph of liquid detection, record chromatograph
Figure, compares gained chromatogram with standard finger-print.
Wherein said standard finger-print is through many batches of qualified samples carry out high performance liquid chromatography detection, records chromatograph
Figure, and the chromatogram of reference object of reference is through statistic computation, generates the standard High Performance liquid chromatogram obtained.
Detection method of the present invention is that unknown sample is carried out high performance liquid chromatography detection, obtains chromatogram, by institute
Obtaining chromatogram and above-mentioned standard finger-print contrasts, similarity is qualified samples higher than 90%, is otherwise defective sample
Product.
For obtaining the standard finger-print of SHENSHUAINING sheet, the present invention works out a kind of efficient liquid phase of SHENSHUAINING sheet further and refers to
Stricture of vagina collection of illustrative plates method for building up, described method comprises the steps:
1) preparation of qualified samples solution:
Take qualified SHENSHUAINING sheet, be ground into powder, take 0.8-1.2g, accurately weighed, to put in tool plug conical flask, precision adds
Enter 45-55% methanol 45-55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, use
45-55% methanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, to obtain final product.
2) preparation of object of reference solution:
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, adds methanol and is respectively prepared every 0.8-1.2ml containing danshensu sodium, berberine hydrochloride
45-55 μ g, containing salvianolic acid B, Hesperidin 80-120 μ g, containing aloe-emodin, rheum emodin, chrysophanol, physcione 8-12 μ g
Solution, to obtain final product.
Separately, Radix Pseudostellariae (0.4-0.6g), Rhizoma Coptidis (0.18-0.22g), the Rhizoma Pinelliae (0.4-0.6g), Pericarpium Citri Reticulatae (0.18-are taken respectively
0.22g), Poria (0.3-0.5g), Radix Et Rhizoma Rhei (0.7-0.9g), Radix Salviae Miltiorrhizae (0.8-1.2g), Radix Achyranthis Bidentatae (0.3-0.5g), Flos Carthami (0.18-
0.22g), Radix Glycyrrhizae (0.18-0.22g) medicinal powder, accurately weighed, put in tool plug conical flask, accurate add 70-90% methanol
20-30ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, supply with 80% methanol and subtract
The weight lost, shakes up, and filters, takes subsequent filtrate, to obtain final product.
3) detection: qualified samples solution and object of reference solution are injected separately into chromatograph of liquid, measures, obtains chromatogram.
4) through multiple batches of qualified samples is measured, after gained chromatogram is comprehensive, standard control fingerprint is finally given
Collection of illustrative plates, has 15 marker peak, and wherein 2, No. 6 peaks are Radix Salviae Miltiorrhizae characteristic peak, and 3,5,9,10,11,12,13,14, No. 15 peaks are Radix Et Rhizoma Rhei
Characteristic peak, No. 4 is Pericarpium Citri Reticulatae characteristic peak, No. 8 for Rhizoma Coptidis characteristic peak.
Preferably, the SHENSHUAINING sheet high-efficiency liquid-phase fingerprint method for building up of the present invention, comprise the steps:
1) preparation of qualified samples solution:
Take qualified SHENSHUAINING sheet, be ground into powder, take 0.8-1.2g, accurately weighed, to put in tool plug conical flask, precision adds
Enter 45-55% methanol 45-55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, use
45-55% methanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, to obtain final product.
2) preparation of object of reference solution:
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, add methanol be respectively prepared every 1ml containing danshensu sodium, berberine hydrochloride 50 μ g, contain
Salvianolic acid B, Hesperidin 100 μ g, containing aloe-emodin, rheum emodin, chrysophanol, the solution of physcione 10 μ g, to obtain final product.
Separately, Radix Pseudostellariae (0.5g), Rhizoma Coptidis (0.2g), the Rhizoma Pinelliae (0.5g), Pericarpium Citri Reticulatae (0.2g), Poria (0.4g), big is taken respectively
The medicinal powders such as yellow (0.8g), Radix Salviae Miltiorrhizae (1.0g), Radix Achyranthis Bidentatae (0.4g), Flos Carthami (0.2g), Radix Glycyrrhizae (0.2g), accurately weighed, put tool
In plug conical flask, accurate addition 80% methanol 25ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30
Minute, take out, let cool, more weighed weight, supply the weight of less loss with 80% methanol, shake up, filter, take subsequent filtrate, to obtain final product.
3) detection: qualified samples solution and object of reference solution are injected separately into chromatograph of liquid, measures, obtains chromatogram.
4) through multiple batches of qualified samples is measured, after gained chromatogram is comprehensive, standard control fingerprint is finally given
Collection of illustrative plates, has 15 marker peak, and wherein 2, No. 6 peaks are Radix Salviae Miltiorrhizae characteristic peak, and 3,5,9,10,11,12,13,14, No. 15 peaks are Radix Et Rhizoma Rhei
Characteristic peak, No. 4 is Pericarpium Citri Reticulatae characteristic peak, No. 8 for Rhizoma Coptidis characteristic peak.
The present invention is to screen relevant detection method on the basis of existing technology, finally gives a kind of practicability and effectiveness
Fingerprint atlas detection method.
The screening technique of the present invention is as follows:
1 instrument and reagent
Instrument: Shimadzu 20A high performance liquid chromatograph (quaternary pump, DAD detector)
Reagent: acetonitrile is chromatographically pure, water is deionized water, and other reagent is analytical pure.
2 chromatographic conditions
Chromatographic column: Agilent Extend C18(4.6 × 250mm, 5 μm)
Flowing phase: with acetonitrile as mobile phase A, the phosphoric acid with 0.1% is as Mobile phase B, and the regulation according to the form below carries out gradient
Eluting；Flow velocity is 1.0ml/min；Detection wavelength is 280nm；Column temperature is 30 DEG C, sample size 10 μ l.
Eluent gradient eluting table
The preparation of 3 need testing solutions takes the sample powder under assay item, takes about 1.0g, accurately weighed, puts tool plug cone
In shape bottle, the accurate 50% methanol 50ml that adds, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 minutes,
Take out, let cool, more weighed weight, supply the weight of less loss with 50% methanol, shake up, filter, take subsequent filtrate, to obtain final product.
4, prepared by contrast solution
4.1 reference substance solution preparations take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, Radix Et Rhizoma Rhei
Element, chrysophanic acid, chrysophanol, physcione reference substance are appropriate, accurately weighed, add methanol be respectively prepared every 1ml containing danshensu sodium,
Berberine hydrochloride 50 μ g, containing salvianolic acid B, Hesperidin 100 μ g, containing aloe-emodin, rheum emodin, chrysophanol, physcione 10 μ
The solution of g, to obtain final product.
4.2 control medicinal material solution preparations take Radix Pseudostellariae (0.5g), Rhizoma Coptidis (0.2g), the Rhizoma Pinelliae (0.5g), Pericarpium Citri Reticulatae respectively
(0.2g), the medicine such as Poria (0.4g), Radix Et Rhizoma Rhei (0.8g), Radix Salviae Miltiorrhizae (1.0g), Radix Achyranthis Bidentatae (0.4g), Flos Carthami (0.2g), Radix Glycyrrhizae (0.2g)
Material powder, accurately weighed, put in tool plug conical flask, accurate addition 80% methanol 25ml, close plug, weighed weight, supersound process (merit
Rate 250W, frequency 40kHz) 30 minutes, take out, let cool, more weighed weight, supply the weight of less loss with 80% methanol, shake up, filter
Cross, take subsequent filtrate, to obtain final product.
The ownership of chromatographic peak
Sample has 15 marker peak, through and control medicinal material comparison, wherein 2, No. 6 peaks are Radix Salviae Miltiorrhizae characteristic peak, 3,5,9,10,
11,12,13,14, No. 15 peaks are Radix Et Rhizoma Rhei characteristic peak, and No. 4 is Pericarpium Citri Reticulatae characteristic peak, No. 8 for Rhizoma Coptidis characteristic peak.
5 methodological studies
5.1 replica tests take same batch sample, prepare 6 parts of need testing solutions simultaneously, and sample introduction measures respectively.Result is each common
Having the relative retention time RSD value at peak between 0.05-0.1%, relative peak area RSD value, between 2.1-3.5%, shows weight
Renaturation is good.
5.2 precision tests take same need testing solution, continuous sample introduction 6 times, measure finger printing.The each total peak of result
Relative retention time RSD value between 0.01～0.2%, relative peak area be RSD value between 1.2～2.1%, show precision
Degree is good.
5.3 stability tests take with a need testing solution, measure at 0,4,8,12,15,24 hours sample introductions respectively, result
The relative retention time RSD value at each total peak is between 0.03-0.4%, and relative peak area value, between 1.1-2.3%, shows
Test sample is at least stable in 24.
The generation of 6 reference fingerprints and Similarity Measure
Take 10 batch samples, be measured by text method, measurement result is imported chromatographic fingerprints of Chinese materia medica similarity and comments
Valency system, generates reference fingerprint.By similarity evaluation system, by the reference fingerprint of test sample finger printing Yu generation
Carrying out Similarity Measure, result see table.
The similarity analysis result of 10 batch samples
Compared to the prior art, accuracy of the present invention is high, highly sensitive, good stability, favorable reproducibility, reality simple to operate
With, achieve unexpected technique effect.
Accompanying drawing explanation
Fig. 1, sample chromatogram figure
Fig. 2, danshensu sodium chromatogram
Fig. 3, salvianolic acid B chromatogram
Fig. 4, Hesperidin chromatogram
Fig. 5, berberine hydrochloride chromatogram
Fig. 6, aloe-emodin chromatogram
Fig. 7, rheum emodin chromatogram
Fig. 8, chrysophanic acid chromatogram
Fig. 9, chrysophanol chromatogram
Figure 10, physcione chromatogram
Figure 11, Pericarpium Citri Reticulatae chromatogram
Figure 12, Radix Et Rhizoma Rhei chromatogram
Figure 13, Radix Salviae Miltiorrhizae chromatogram
Figure 14, Rhizoma Coptidis chromatogram
Figure 15, Radix Glycyrrhizae chromatogram
Figure 16, Rhizoma Pinelliae Preparatum chromatogram
Figure 17, Poria chromatogram
Figure 18, Flos Carthami chromatogram
Figure 19, Radix Achyranthis Bidentatae chromatogram
Figure 20, Radix Pseudostellariae chromatogram
Figure 21, standard finger-print
Figure 22,10 batch sample similarity collection of illustrative plates
Detailed description of the invention
Further illustrate the present invention by the following examples.
Embodiment 1
Step 1: prepared by reference substance solution
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, adds methanol and is respectively prepared every 0.8-1.2ml containing danshensu sodium, berberine hydrochloride
45-55 μ g, containing salvianolic acid B, Hesperidin 80-120 μ g, containing aloe-emodin, rheum emodin, chrysophanol, physcione 8-12 μ g
Solution, to obtain final product.
Step 2: prepared by control medicinal material solution
Take Radix Pseudostellariae (0.4-0.6g), Rhizoma Coptidis (0.18-0.22g), the Rhizoma Pinelliae (0.4-0.6g), Pericarpium Citri Reticulatae (0.18-respectively
0.22g), Poria (0.3-0.5g), Radix Et Rhizoma Rhei (0.7-0.9g), Radix Salviae Miltiorrhizae (0.8-1.2g), Radix Achyranthis Bidentatae (0.3-0.5g), Flos Carthami (0.18-
0.22g), Radix Glycyrrhizae (0.18-0.22g) medicinal powder, accurately weighed, put in tool plug conical flask, accurate add 70-90% methanol
20-30ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, supply with 80% methanol and subtract
The weight lost, shakes up, and filters, takes subsequent filtrate, to obtain final product.
Step 3: prepared by need testing solution
Take sample powder 0.8-1.2g, accurately weighed, put in tool plug conical flask, accurate addition 45-55% methanol 45-
55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, supply less loss with 50% methanol
Weight, shake up, filter, take subsequent filtrate, to obtain final product.
Step 4: algoscopy
Take reference substance solution and need testing solution each 5-15 μ L respectively, be injected separately into chromatograph of liquid detection, record chromatograph
Figure, compares gained chromatogram with standard finger-print.
Preferably, the detection method of the present invention, comprise the steps:
Step 1: prepared by reference substance solution
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, add methanol be respectively prepared every 1ml containing danshensu sodium, berberine hydrochloride 50 μ g, contain
Salvianolic acid B, Hesperidin 100 μ g, containing aloe-emodin, rheum emodin, chrysophanol, the solution of physcione 10 μ g, to obtain final product.
Step 2: prepared by control medicinal material solution
Take Radix Pseudostellariae (0.5g), Rhizoma Coptidis (0.2g), the Rhizoma Pinelliae (0.5g), Pericarpium Citri Reticulatae (0.2g), Poria (0.4g), Radix Et Rhizoma Rhei respectively
(0.8g), Radix Salviae Miltiorrhizae (1.0g), Radix Achyranthis Bidentatae (0.4g), Flos Carthami (0.2g), the medicinal powder such as Radix Glycyrrhizae (0.2g), accurately weighed, put tool plug
In conical flask, accurate addition 80% methanol 25ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 points
Clock, takes out, lets cool, more weighed weight, supplies the weight of less loss with 80% methanol, shakes up, and filters, takes subsequent filtrate, to obtain final product.
Step 3: prepared by need testing solution
Take the sample powder under assay item, take about 1.0g, accurately weighed, put in tool plug conical flask, accurate addition
50% methanol 50ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 minutes, take out, let cool, then claim
Determine weight, supply the weight of less loss with 50% methanol, shake up, filter, take subsequent filtrate, to obtain final product.
Step 4: algoscopy
Take reference substance solution and need testing solution each 1-6 μ L respectively, be injected separately into chromatograph of liquid detection, record chromatograph
Figure, compares gained chromatogram with standard finger-print.
The present invention open a kind of SHENSHUAINING sheet high-efficiency liquid-phase fingerprint method for building up further, described method includes as follows
Step:
1) preparation of qualified samples solution:
Take qualified SHENSHUAINING sheet, be ground into powder, take 0.8-1.2g, accurately weighed, to put in tool plug conical flask, precision adds
Enter 45-55% methanol 45-55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, use
45-55% methanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, to obtain final product.
2) preparation of object of reference solution:
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, adds methanol and is respectively prepared every 0.8-1.2ml containing danshensu sodium, berberine hydrochloride
45-55 μ g, containing salvianolic acid B, Hesperidin 80-120 μ g, containing aloe-emodin, rheum emodin, chrysophanol, physcione 8-12 μ g
Solution, to obtain final product.
Separately, Radix Pseudostellariae (0.4-0.6g), Rhizoma Coptidis (0.18-0.22g), the Rhizoma Pinelliae (0.4-0.6g), Pericarpium Citri Reticulatae (0.18-are taken respectively
0.22g), Poria (0.3-0.5g), Radix Et Rhizoma Rhei (0.7-0.9g), Radix Salviae Miltiorrhizae (0.8-1.2g), Radix Achyranthis Bidentatae (0.3-0.5g), Flos Carthami (0.18-
0.22g), Radix Glycyrrhizae (0.18-0.22g) medicinal powder, accurately weighed, put in tool plug conical flask, accurate add 70-90% methanol
20-30ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, supply with 80% methanol and subtract
The weight lost, shakes up, and filters, takes subsequent filtrate, to obtain final product.
3) detection: qualified samples solution and object of reference solution are injected separately into chromatograph of liquid, measures, obtains chromatogram.
4) through multiple batches of qualified samples is measured, after gained chromatogram is comprehensive, standard control fingerprint is finally given
Collection of illustrative plates, has 15 marker peak, and wherein 2, No. 6 peaks are Radix Salviae Miltiorrhizae characteristic peak, and 3,5,9,10,11,12,13,14, No. 15 peaks are Radix Et Rhizoma Rhei
Characteristic peak, No. 4 is Pericarpium Citri Reticulatae characteristic peak, No. 8 for Rhizoma Coptidis characteristic peak.
Preferably, the SHENSHUAINING sheet high-efficiency liquid-phase fingerprint method for building up of the present invention, comprise the steps:
1) preparation of qualified samples solution:
Take qualified SHENSHUAINING sheet, be ground into powder, take 0.8-1.2g, accurately weighed, to put in tool plug conical flask, precision adds
Enter 45-55% methanol 45-55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, use
45-55% methanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, to obtain final product.
2) preparation of object of reference solution:
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol,
Physcione reference substance is appropriate, accurately weighed, add methanol be respectively prepared every 1ml containing danshensu sodium, berberine hydrochloride 50 μ g, contain
Salvianolic acid B, Hesperidin 100 μ g, containing aloe-emodin, rheum emodin, chrysophanol, the solution of physcione 10 μ g, to obtain final product.
Separately, Radix Pseudostellariae (0.5g), Rhizoma Coptidis (0.2g), the Rhizoma Pinelliae (0.5g), Pericarpium Citri Reticulatae (0.2g), Poria (0.4g), big is taken respectively
The medicinal powders such as yellow (0.8g), Radix Salviae Miltiorrhizae (1.0g), Radix Achyranthis Bidentatae (0.4g), Flos Carthami (0.2g), Radix Glycyrrhizae (0.2g), accurately weighed, put tool
In plug conical flask, accurate addition 80% methanol 25ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30
Minute, take out, let cool, more weighed weight, supply the weight of less loss with 80% methanol, shake up, filter, take subsequent filtrate, to obtain final product.
3) detection: qualified samples solution and object of reference solution are injected separately into chromatograph of liquid, measures, obtains chromatogram.
4) through multiple batches of qualified samples is measured, after gained chromatogram is comprehensive, standard control fingerprint is finally given
Collection of illustrative plates, has 15 marker peak, and wherein 2, No. 6 peaks are Radix Salviae Miltiorrhizae characteristic peak, and 3,5,9,10,11,12,13,14, No. 15 peaks are Radix Et Rhizoma Rhei
Characteristic peak, No. 4 is Pericarpium Citri Reticulatae characteristic peak, No. 8 for Rhizoma Coptidis characteristic peak.
Claims (4)
1. the detection method of a SHENSHUAINING sheet, it is characterised in that described method comprises the steps:
Step 1: prepared by reference substance solution
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol, Radix Et Rhizoma Rhei
Element methyl ether reference substance is appropriate, accurately weighed, adds methanol and is respectively prepared every 0.8-1.2ml containing danshensu sodium, berberine hydrochloride 45-55
μ g, containing salvianolic acid B, Hesperidin 80-120 μ g, containing aloe-emodin, rheum emodin, chrysophanol, physcione 8-12 μ g molten
Liquid, to obtain final product；
Step 2: prepared by control medicinal material solution
Take respectively Radix Pseudostellariae (0.4-0.6g), Rhizoma Coptidis (0.18-0.22g), the Rhizoma Pinelliae (0.4-0.6g), Pericarpium Citri Reticulatae (0.18-0.22g),
Poria (0.3-0.5g), Radix Et Rhizoma Rhei (0.7-0.9g), Radix Salviae Miltiorrhizae (0.8-1.2g), Radix Achyranthis Bidentatae (0.3-0.5g), Flos Carthami (0.18-0.22g),
Radix Glycyrrhizae (0.18-0.22g) medicinal powder, accurately weighed, put in tool plug conical flask, accurate addition 70-90% methanol 20-30ml,
Close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, the weight of less loss is supplied with 80% methanol
Amount, shakes up, and filters, takes subsequent filtrate, to obtain final product；
Step 3: prepared by need testing solution
Take sample powder 0.8-1.2g, accurately weighed, put in tool plug conical flask, the accurate 45-55% methanol 45-55ml that adds, close
Plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, the weight of less loss is supplied with 50% methanol,
Shake up, filter, take subsequent filtrate, to obtain final product；
Step 4: algoscopy
Take contrast solution and need testing solution each 5-15 μ L respectively, be injected separately into chromatograph of liquid detection, record chromatogram, by institute
Obtain chromatogram to compare with standard finger-print.
Detection method the most according to claim 1, it is characterised in that comprise the steps:
Step 1: prepared by reference substance solution
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol, Radix Et Rhizoma Rhei
Element methyl ether reference substance is appropriate, accurately weighed, add methanol be respectively prepared every 1ml containing danshensu sodium, berberine hydrochloride 50 μ g, containing red phenol
Acid B, Hesperidin 100 μ g, containing aloe-emodin, rheum emodin, chrysophanol, the solution of physcione 10 μ g, to obtain final product；
Step 2: prepared by control medicinal material solution
Take Radix Pseudostellariae (0.5g), Rhizoma Coptidis (0.2g), the Rhizoma Pinelliae (0.5g), Pericarpium Citri Reticulatae (0.2g), Poria (0.4g), Radix Et Rhizoma Rhei respectively
(0.8g), Radix Salviae Miltiorrhizae (1.0g), Radix Achyranthis Bidentatae (0.4g), Flos Carthami (0.2g), the medicinal powder such as Radix Glycyrrhizae (0.2g), accurately weighed, put tool plug
In conical flask, accurate addition 80% methanol 25ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 points
Clock, takes out, lets cool, more weighed weight, supplies the weight of less loss with 80% methanol, shakes up, and filters, takes subsequent filtrate, to obtain final product；
Step 3: prepared by need testing solution
Take the sample powder under assay item, take about 1.0g, accurately weighed, put in tool plug conical flask, accurate addition 50% first
Alcohol 50ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 minutes, take out, let cool, more weighed weight,
Supply the weight of less loss with 50% methanol, shake up, filter, take subsequent filtrate, to obtain final product；
Step 4: algoscopy
Take contrast solution and need testing solution each 1-6 μ L respectively, be injected separately into chromatograph of liquid detection, record chromatogram, by institute
Obtain chromatogram to compare with standard finger-print.
3. a SHENSHUAINING sheet high-efficiency liquid-phase fingerprint method for building up, it is characterised in that described method comprises the steps:
1) preparation of qualified samples solution:
Take qualified SHENSHUAINING sheet, be ground into powder, take 0.8-1.2g, accurately weighed, put in tool plug conical flask, accurate addition
45-55% methanol 45-55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, use 45-
55% methanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, to obtain final product；
2) preparation of object of reference solution:
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol, Radix Et Rhizoma Rhei
Element methyl ether reference substance is appropriate, accurately weighed, adds methanol and is respectively prepared every 0.8-1.2ml containing danshensu sodium, berberine hydrochloride 45-55
μ g, containing salvianolic acid B, Hesperidin 80-120 μ g, containing aloe-emodin, rheum emodin, chrysophanol, physcione 8-12 μ g molten
Liquid, to obtain final product；
Separately, Radix Pseudostellariae (0.4-0.6g), Rhizoma Coptidis (0.18-0.22g), the Rhizoma Pinelliae (0.4-0.6g), Pericarpium Citri Reticulatae (0.18-are taken respectively
0.22g), Poria (0.3-0.5g), Radix Et Rhizoma Rhei (0.7-0.9g), Radix Salviae Miltiorrhizae (0.8-1.2g), Radix Achyranthis Bidentatae (0.3-0.5g), Flos Carthami (0.18-
0.22g), Radix Glycyrrhizae (0.18-0.22g) medicinal powder, accurately weighed, put in tool plug conical flask, accurate add 70-90% methanol
20-30ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, supply with 80% methanol and subtract
The weight lost, shakes up, and filters, takes subsequent filtrate, to obtain final product；
3) detection: qualified samples solution and object of reference solution are injected separately into chromatograph of liquid, measures, obtains chromatogram；
4) through multiple batches of qualified samples is measured, after gained chromatogram is comprehensive, standard control finger printing is finally given,
Having 15 marker peak, wherein 2, No. 6 peaks are Radix Salviae Miltiorrhizae characteristic peak, and 3,5,9,10,11,12,13,14, No. 15 peaks are Radix Et Rhizoma Rhei feature
Peak, No. 4 is Pericarpium Citri Reticulatae characteristic peak, No. 8 for Rhizoma Coptidis characteristic peak.
Fingerprint the most according to claim 3, it is characterised in that comprise the steps:
1) preparation of qualified samples solution:
Take qualified SHENSHUAINING sheet, be ground into powder, take 0.8-1.2g, accurately weighed, put in tool plug conical flask, accurate addition
45-55% methanol 45-55ml, close plug, weighed weight, supersound process 25-35 minute, take out, let cool, more weighed weight, use 45-
55% methanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, to obtain final product；
2) preparation of object of reference solution:
Take danshensu sodium, salvianolic acid B, Hesperidin, berberine hydrochloride, aloe-emodin, rheum emodin, chrysophanic acid, chrysophanol, Radix Et Rhizoma Rhei
Element methyl ether reference substance is appropriate, accurately weighed, add methanol be respectively prepared every 1ml containing danshensu sodium, berberine hydrochloride 50 μ g, containing red phenol
Acid B, Hesperidin 100 μ g, containing aloe-emodin, rheum emodin, chrysophanol, the solution of physcione 10 μ g, to obtain final product；
Separately, Radix Pseudostellariae (0.5g), Rhizoma Coptidis (0.2g), the Rhizoma Pinelliae (0.5g), Pericarpium Citri Reticulatae (0.2g), Poria (0.4g), Radix Et Rhizoma Rhei are taken respectively
(0.8g), Radix Salviae Miltiorrhizae (1.0g), Radix Achyranthis Bidentatae (0.4g), Flos Carthami (0.2g), the medicinal powder such as Radix Glycyrrhizae (0.2g), accurately weighed, put tool plug
In conical flask, accurate addition 80% methanol 25ml, close plug, weighed weight, supersound process (power 250W, frequency 40kHz) 30 points
Clock, takes out, lets cool, more weighed weight, supplies the weight of less loss with 80% methanol, shakes up, and filters, takes subsequent filtrate, to obtain final product；
3) detection: qualified samples solution and object of reference solution are injected separately into chromatograph of liquid, measures, obtains chromatogram；
4) through multiple batches of qualified samples is measured, after gained chromatogram is comprehensive, standard control finger printing is finally given,
Having 15 marker peak, wherein 2, No. 6 peaks are Radix Salviae Miltiorrhizae characteristic peak, and 3,5,9,10,11,12,13,14, No. 15 peaks are Radix Et Rhizoma Rhei feature
Peak, No. 4 is Pericarpium Citri Reticulatae characteristic peak, No. 8 for Rhizoma Coptidis characteristic peak.
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| CN109752472B (en) * | 2019-01-11 | 2022-01-28 | 吉林省现代中药工程研究中心有限公司 | Method for constructing characteristic spectrum of compound rohdea japonica capsules and method for detecting quality of compound rohdea japonica capsules |
| CN109900832A (en) * | 2019-04-17 | 2019-06-18 | 南京海昌中药集团有限公司 | A kind of detection method of rhizoma pinellinae praeparata finger-print |
| CN110927302A (en) * | 2019-12-08 | 2020-03-27 | 山东沃华医药科技股份有限公司 | Method for measuring fingerprint of collateral-dredging phlegm-reducing capsule |
| CN110927302B (en) * | 2019-12-08 | 2022-04-22 | 山东沃华医药科技股份有限公司 | Method for measuring fingerprint of collateral-dredging phlegm-reducing capsule |
| CN114636760A (en) * | 2020-12-15 | 2022-06-17 | 云南雷允上理想药业有限公司 | Fingerprint spectrum establishment method and application of traditional Chinese medicine composition for treating kidney diseases |
| CN114636760B (en) * | 2020-12-15 | 2024-01-02 | 云南雷允上理想药业有限公司 | Fingerprint spectrum establishment method of traditional Chinese medicine composition for treating kidney diseases and application of fingerprint spectrum establishment method |
| WO2023024322A1 (en) * | 2021-08-24 | 2023-03-02 | 石家庄以岭药业股份有限公司 | Method for determining fingerprint of traditional chinese medicine composition |
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Denomination of invention: A Method for Determining the Effective Components of Shenshuening Tablets Effective date of registration: 20230925 Granted publication date: 20180626 Pledgee: Bank of China Limited Qinhuangdao Branch Pledgor: QINHUANGDAO SHANHAIGUAN PHARMACEUTICAL Co.,Ltd. Registration number: Y2023980058918 |