CN107589185A - Train of thought leads to the fingerprint atlas detection method and its finger-print of particle - Google Patents
Train of thought leads to the fingerprint atlas detection method and its finger-print of particle Download PDFInfo
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Abstract
The invention discloses a kind of train of thought to lead to particle fingerprint atlas detection method, and this method is established using high performance liquid chromatography, and chromatographic condition includes：Using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase A, using 0.3 volume % aqueous formic acids as Mobile phase B, gradient elution, 0.5~1.5mlmin of flow velocity 1,20~35 DEG C of column temperature, 210~360nm of Detection wavelength；Theoretical cam curve is calculated by rutin peak should be not less than 3000.Experimental verification shows, the detection method of the present invention, stability and it is reproducible, analyze speed is fast, analysis efficiency is high, avoid the unicity and one-sidedness of prior art quality control, so as to quality that is objective, comprehensively and accurately evaluating the logical particle of train of thought, the quality of particle is led to control train of thought and ensures that clinical efficacy is significant.On the other hand, present invention also offers the train of thought obtained by the logical particle fingerprint atlas detection method of train of thought as described above to lead to particle finger-print.
Description
Technical field
The invention belongs to Chinese medicine detection technique field, specifically, is related to the finger-print detection that a kind of train of thought leads to particle
Method and its finger-print.
Background technology
Train of thought, which leads to particle, has qi and activate blood circulation, blood-activating analgetic, active function of promoting blood circulation.Suitable for the heart caused by the treatment obstruction of qi in the chest
Chest pain, shortness of breath uncomfortable in chest, dizziness of having a headache and coronary heart diseases and angina pectoris have above-mentioned all diseases, and extremity numbness, half body caused by apoplexy are not
Then disease is waited.
Train of thought is led to particle prescription and come from《Chinese medicine ministry standard》" train of thought leads to particle ", and produced without competition in 1987, on
City.Train of thought is led in the prescription of particle, with Radix Codonopsis, when being classified as monarch drug in a prescription, QI invigorating and with blood-nourishing；Safflower, the red sage root, Ligusticum wallichii, earthworm are minister
Medicine, auxiliary monarch drug in a prescription the effect of reaching blood circulation and channel invigorating；Hawthorn is adjutant, and pass through qi and blood, promoting blood circulation analgesic；Scouring rush, the root of kudzu vine, the sophora bud
To make medicine, clear and coherent numbness can be risen, helps medicine up, guides drugs to illness station.Monarch, complete square compatibility 5, qi and activate blood circulation, blood-activating analgetic, fit
Treating Patients of Angina Pectoris for qi deficiency to blood stasis, heart vessel blockage stasis.
After traditional Chinese medicine fingerprint refers to that some Chinese medicines or Chinese medicine preparation are appropriately processed, using certain analysis means,
What is obtained can indicate the chromatogram or spectrogram of its chemical feature.Traditional Chinese medicine fingerprint is a kind of synthesis, quantifiable mirror
Determine means, it is built upon on the basis of chemical composition of Chinese materia medica system research, is mainly used in evaluating Chinese medicine and Chinese medicine preparation
Authenticity, Optimality and the stability of semi-manufactured goods quality." globality " and " ambiguity " is its distinguishing feature.At this stage, in
The effective elements of the medicine is most without in the case of clearly, and traditional Chinese medicine fingerprint is for effectively control Chinese medicine or Chinese patent drug
Quality has great importance.The main manufacturing enterprise of Japanese Kampo medicine is in the 1980s just in enterprises using high
Imitate liquid-phase fingerprint control quality.Germany and France also with high-efficiency liquid-phase fingerprint method to ginkgo biloba p.e
Carry out quality control, U.S. recent years the also method of quality control clearly using finger-print as mixture group.With research
Go deep into, it has been found that, as the product of putting into practice of theory of traditional Chinese medical science, Chinese medicine, especially herbal mixture, it is contained therein it is any into
Its overall curative effect can not all be represented by dividing.People gradually recognize that the quality standard of existing reference chemical drug quality control model is not
In reflection Chinese medicine that can be appropriate quality.From existing quality control model to a kind of synthesis, macroscopical, quantifiable mirror
It has been the trend developed not combined with principle active component assay.
Train of thought leads to particle and shares 10 taste medicinal materials, and it is not only to carry out assay to principal component Puerarin in existing quality standard
No more, it is necessary to its material group is integrally controlled.So in addition to " micro-analysis ", certain " macroscopic view point should be also used
Analysis " method, effectively characterizes traditional Chinese medicine quality on the whole.So establish the fingerprint atlas detection method gesture that a kind of train of thought leads to particle
It must go.
The content of the invention
Inventor developed the fingerprint atlas detection method and its finger-print that a kind of train of thought leads to particle, pass through the method
It can ensure that train of thought leads to quality stability, uniformity and the controllability of particle, so that it is guaranteed that train of thought is led to the security of particle and had
Effect property.
It is an object of the invention to provide the fingerprint atlas detection method that a kind of train of thought leads to particle.
The second object of the present invention is to provide the finger-print for the fingerprint atlas detection method for leading to particle for train of thought.
In embodiments of the invention, the invention provides the fingerprint atlas detection method that a kind of train of thought leads to particle, bag
Include and particle is led to train of thought using high performance liquid chromatography detected, wherein, chromatographic condition includes：
Using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase A, with 0.3 volume % aqueous formic acids
For Mobile phase B, gradient elution, 0.5~1.5mlmin of flow velocity-1, 20~35 DEG C of column temperature, 210~360nm of Detection wavelength；It is theoretical
The number of plates is calculated by rutin peak should be not less than 3000.
In a preferred embodiment of the invention, a kind of train of thought provided by the invention leads to the finger-print detection side of particle
Method, wherein, during gradient elution, the ratio of mobile phase A, which becomes, to be turned to：0~3min, 5 volume % acetonitriles；3~50min, 5 bodies
The product volume % acetonitriles of %~42；The volume % acetonitriles of 50~50.1min, 42 volume %~95；50.1~60min, 95 volume % second
Nitrile；That is gradient elution program such as following table：
In a preferred embodiment of the invention, a kind of train of thought provided by the invention leads to the finger-print detection side of particle
Method, wherein, the chromatographic column using octadecylsilane chemically bonded silica as filler is phenomenenx luna (2) C18
(4.6*250mm, 5um).
In a preferred embodiment of the invention, a kind of train of thought provided by the invention leads to the finger-print detection side of particle
Method, it is further comprising the steps of：
(1) preparation of need testing solution：Train of thought is taken to lead to particle 2.0g, it is accurately weighed, put in 100ml conical flask with cover, essence
70 volume 25~75ml of % ethanol of close addition, close plug, weighed weight are ultrasonically treated (250w, 40KHz) 10~40 minutes, let cool,
The weight of less loss is supplied with 70 volume % ethanol, is shaken up, is filtered, is taken subsequent filtrate, produce；
(2) preparation of reference substance solution：Take Puerarin, rutin, forulic acid, lobetyolin, tanshin polyphenolic acid B, Quercetin, Kaempferia galanga
It is plain appropriate, add 70 volume % ethanol that every 1ml is made containing μ g of Puerarin 112, μ g of rutin 204, μ g of forulic acid 44, the μ of lobetyolin 32
G, μ g of tanshin polyphenolic acid B 92, μ g of Quercetin 40, the μ g of Kaempferide 28 mixed reference substance solution, are produced；
(3) determine：It is accurate respectively to draw reference substance solution and each 2~10 μ l of need testing solution, inject high performance liquid chromatography
Instrument, the chromatogram in 60 minutes is recorded, used《Similarity evaluation》2004A versions to collection of illustrative plates at
Reason, produce the finger-print that train of thought leads to particle.
In the more preferred of the present invention, the invention provides the finger-print detection side that a kind of train of thought leads to particle
Method, comprise the following steps：
(1) preparation of need testing solution：Train of thought is taken to lead to particle 2.0g, it is accurately weighed, put in 100ml conical flask with cover, essence
70 volume % ethanol 50ml of close addition, close plug, weighed weight, ultrasonic (250w, 40KHz) are handled 20 minutes, let cool, with 70 bodies
Product % ethanol supplies the weight of less loss, shakes up, and filters, takes subsequent filtrate, produce；
(2) preparation of reference substance solution：Take Puerarin, rutin, forulic acid, lobetyolin, tanshin polyphenolic acid B, Quercetin, Kaempferia galanga
It is plain appropriate, add 70 volume % ethanol that every 1ml is made containing μ g of Puerarin 112, μ g of rutin 204, μ g of forulic acid 44, the μ of lobetyolin 32
G, μ g of tanshin polyphenolic acid B 92, μ g of Quercetin 40, the μ g of Kaempferide 28 mixed reference substance solution, are produced；
(3) determine：It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, inject high performance liquid chromatograph, note
The chromatogram in 60 minutes is recorded, is used《Similarity evaluation》2004A versions are handled collection of illustrative plates, i.e.,
Obtain the finger-print that train of thought leads to particle；
Here, the chromatographic condition of the high performance liquid chromatography measure is：Using octadecylsilane chemically bonded silica as filler；
Using acetonitrile as mobile phase A；Using 0.3% aqueous formic acid as Mobile phase B；Gradient elution, flow velocity 1.0mlmin-1；25 DEG C of column temperature；
Detection wavelength 270nm；Theoretical cam curve is calculated by rutin first peak should be not less than 3000.
On the other hand, the train of thought obtained the invention provides the fingerprint atlas detection method for leading to particle by above-mentioned train of thought is led to
Particle finger-print, there are 21 shared peaks in the collection of illustrative plates.
In a preferred embodiment of the invention, train of thought provided by the invention leads to particle finger-print, has in the collection of illustrative plates
21 shared peaks, wherein, the retention time at each shared peak is respectively：Shared peak 1：13.234min；Shared peak 2：15.952min；
Shared peak 3：17.602min；Shared peak 4：20.612min；Shared peak 5：21.099min；Shared peak 6：21.321min；It is shared
Peak 7：24.057min；Shared peak 8：27.190min；Shared peak 9：28.845min；Shared peak 10：29.452min；Shared peak
11：29.830min；Shared peak 12：30.206min；Shared peak 13：33.715min；Shared peak 14：34.979min；Shared peak
15：37.136min；Shared peak 16：38.789min；Shared peak 17：39.278min；Shared peak 18：39.610min；Shared peak
19：41.671min；Shared peak 20：45.049min；Shared peak 21：48.173min.
The beneficial effects of the invention are as follows：
(1) train of thought established with method provided by the present invention leads to particle finger-print, can effectively characterize train of thought and lead to
The quality of particle, be advantageous to the quality of overall monitor medicine.
(2) finger-print focuses on each tandem and correlation for forming fingerprint characteristic peak, and it is special to focus on overall looks
Sign, the one-sidedness for judging that train of thought leads to granular mass because determining individual chemical composition was both avoided, had been reduced again for requisite quality
And the possibility artificially handled.
(3) the inventive method have method it is easy, stably, the advantages of precision is high, favorable reproducibility.
In embodiments of the invention, it is related to " % ", unless otherwise specified, all referring to percent by volume.
Brief description of the drawings
Fig. 1 is that 14 batches of train of thoughts that the present invention measures lead to particle finger-print stacking chart.
Fig. 2 is the control collection of illustrative plates that the logical particle finger-print of train of thought is measured by the present invention.
Fig. 3 is that the train of thought that the present invention measures leads to particle reference substance solution collection of illustrative plates, and abscissa is the time (min) in figure, indulges and sits
It is designated as absorbance (mAU).
Fig. 4 is to lead to particle typical case's need testing solution finger-print by the train of thought that measures of the present invention, when abscissa is in figure
Between (min), ordinate is absorbance (mAU).
Embodiment
Comparative example
Train of thought is led in the existing quality standard of particle only controls drug quality by this index of Puerarin, has certain
One-sidedness；And the train of thought of the present invention leads to particle fingerprint atlas detection method and can pass through the finger-print measured with compareing collection of illustrative plates
Similarity software carries out the complete contrast at 21 shared peaks, more comprehensively can accurately characterize the quality of medicine.According to historical data
Show, train of thought leads in particle that puerarin content is in 4.5mg or so, and from following table, puerarin content is near 4.5mg
The similarity of batch medicine finger-print is smaller, traces it to its cause and finds reed in 16110281 and 16101141 liang of batch finger-prints
Ding Fengyu control collection of illustrative plates differences are larger, and tanshin polyphenolic acid B peak differs larger with control collection of illustrative plates in 16102381 batch finger-prints,
Quercetin peak differs larger with control collection of illustrative plates in 16101041 batch finger-prints, causes not high with the similarity for compareing collection of illustrative plates.
And content it is relatively low 2 batch medicine fingerprint similarities it is higher, be found that while that puerarin peak is slightly smaller by comparative analysis, but
Remaining 20 peak approaches with compareing collection of illustrative plates, so with compareing collection of illustrative plates similarity height.Briefly, train of thought, which leads to particle finger-print, is
The quality of medicine is embodied by contrasting 21 peaks, it is more comprehensively accurate to be controlled than single index Puerarin.
The train of thought of table 1 leads to granular mass control contrast
Embodiment
Train of thought leads to the construction method of particle finger-print, leads to particle to train of thought using liquid chromatography and is measured, specifically
Method is as follows：
(1) preparation of reference substance solution：
Puerarin, rutin, forulic acid, lobetyolin, tanshin polyphenolic acid B, Quercetin, appropriate Kaempferide are taken, adds 70% volume second
Every 1ml is made containing μ g of Puerarin 112, μ g of rutin 204, μ g of forulic acid 44, μ g of lobetyolin 32, μ g of tanshin polyphenolic acid B 92, quercitrin in alcohol
40 μ g of element, the μ g of Kaempferide 28 mixed reference substance solution, are produced.
(2) preparation of need testing solution：
Train of thought is taken to lead to particle 2.0g, it is accurately weighed, put in 100ml conical flask with cover, precision adds 70 volume % ethanol
50ml, close plug, weighed weight, ultrasonic (250w, 40KHz) are handled 20 minutes, are let cool, the weight of less loss is supplied with 70 volume % ethanol
Amount, shakes up, and filters, takes subsequent filtrate, produce.
(3) chromatographic condition and system suitability
Chromatographic column：Phenomenenx luna (2) C18 (4.6*250mm, 5um)；
Mobile phase：A pumps：Acetonitrile；B pumps：0.3 volume % formic acid；
Gradient elution program：
| Time (min) | A (acetonitrile) volumes % | B (0.30 volume % formic acid) volume % |
| 0 | 5 | 95 |
| 3 | 5 | 95 |
| 50 | 42 | 58 |
| 50.1 | 95 | 5 |
| 60 | 95 | 5 |
Detection wavelength：270nm；
Flow velocity：1.0ml；
Column temperature：25℃
Theoretical cam curve is not less than 3000 based on rutin.
(4) determine：It is accurate respectively to draw reference substance solution and each 5 μ l of need testing solution, inject high performance liquid chromatograph, note
The chromatogram in 60 minutes is recorded, is used《Similarity evaluation》2004A versions are handled collection of illustrative plates, i.e.,
Obtain the finger-print that train of thought leads to particle.See Fig. 1-2.
Liquid chromatograph is the Infinity Series high performance liquid chromatographs of Agilent 1260.
There are 21 shared peaks in finger-print, by being detected to reference substance solution and need testing solution simultaneously, identify it
In No. 4 peaks be Puerarin, No. 8 peaks are rutin, No. 9 peaks are forulic acid, No. 13 peaks are lobetyolin, No. 15 peaks are tanshin polyphenolic acid B, 19
Number peak is Quercetin, No. 21 peaks are Kaempferide.Detected additionally by LC-MS, deduce 12 compounds, respectively No. 1 peak again
Codopiloic acid, No. 2 peak 6- hydroxyl Kaempferol 3- rutinose -6- glucosides, No. 3 peak genistins, No. 5 peak Puerarin xylosides, 6
Number peak 3/- methoxy puerarins, No. 7 peak daidzins, No. 10 peak tangshenoside I, No. 11 peak carthamus tinctorius yellow colour As, No. 16 peak daidzins
Member, No. 17 peak salviandic acid As, No. 18 peak Biochanin As, No. 20 peak salvianolic acid Cs.See Fig. 3-4.
Test sample preparation method is investigated
(1) investigation of Extraction solvent
Investigated respectively 10% methanol, 30% methanol, 50% methanol, 70% methanol, methanol, 10% ethanol, 30% ethanol,
50% ethanol, 70% ethanol, ethanol, as a result 70% ethanol and methanol extraction effect are suitable, from environmentally friendly and cost-effective consideration, choosing
Select the Extraction solvent that 70% ethanol leads to particle as train of thought.
(2) investigation of different solvents volume
The Extraction solvent to tri- kinds of different volumes of 25ml, 50ml, 75ml is investigated that (corresponding sample size is respectively
2.5ul, 5ul, 7.5ul), sample peak area is generally more than sample when Extraction solvent is 25ml when as a result Extraction solvent is 50ml
Peak area, but 75ml Extraction solvents relatively have no advantage with 50ml, select the ethanol of 50ml 70% to lead to the extraction of particle as train of thought
Solvent volume.
(3) investigation of different extracting modes
Two kinds of extracting modes of refluxing extraction and ultrasonic extraction are compared, as a result both displays are without significant difference, selection ultrasound
Extract the extracting mode for leading to particle as train of thought.
(4) investigation of different extraction times
The extraction time that 5min, 10min, 20min, 30min, 40min lead to particulate samples as train of thought has been investigated respectively, knot
Three's peak area is more or less the same when fruit extraction time is 20min, 30min, 40min, but more with the obvious advantage than 5min, 10min, selection
20min is the ultrasonic extraction time that train of thought leads to particle.
Train of thought leads to the investigation of particulate chromatography condition
(1) investigation of elution system
4 kinds of acetonitrile-water, acetonitrile-sour water, methanol-water, methanol-sour water different types of elution systems are compared for respectively, are tied
Appearance is very fast when fruit organic phase is acetonitrile, and most materials can be eluted in 50min, special when adding sour in mobile phase
Levy peak forulic acid and tanshin polyphenolic acid B appearance time in advance and peak type is preferable, therefore select elution system of the acetonitrile-sour water as this product.
(2) investigation of variety classes acid
It compared for the formic acid of acetonitrile -0.3%, species is often used in 3 kinds of acetic acid of acetonitrile -0.5%, the phosphoric acid of acetonitrile -0.05% laboratories
And the acid of concentration is compared, it is found that the formic acid of acetonitrile -0.3%, the phosphoric acid of acetonitrile -0.05% separating effect will be compared with the second of acetonitrile -0.5%
Acid is good, it is also contemplated that phosphoric acid is unable to feed liquor matter combined system, final choice formic acid leads to the PH regulations of particle flow phase as train of thought
Agent.
(3) investigation of different acid concentrations
0.2% formic acid, 0.3% formic acid, 0.4% formic acid are compared for respectively, and as a result 0.3% formic acid separating effect is best, institute
Lead to the mobile phase of particle using the formic acid of final choice acetonitrile -0.3% as train of thought.
(4) investigation of different chromatographic columns
Phenomenenx luna (2) C18 (4.6*250mm, 5um), Agilent plus C18 (4.6* are compared for respectively
250mm, 5um), the C18 chromatographic columns of 3 kinds of different brands of waters Atlantis T3C18 (4.6*250mm, 5um), as a result only
There are phenomenenx luna (2) C18 chromatographic columns can be by Puerarin xyloside and 3/- methoxy puerarin is separated, other 2 sections of colors
Compose post not separate completely, therefore select phenomenenx luna (2) C18 (4.6*250mm, 5um) to lead to particle fingerprint as train of thought
Chromatographic column during atlas analysis.
(5) investigation of different column temperatures
20 DEG C, 25 DEG C, 30 DEG C of 3 kinds of column temperatures are compared for, as a result 25 DEG C of separating effects are best, therefore select 25 DEG C to lead to as train of thought
Column temperature during particle fingerprint map analyzing.
(6) investigation of different wave length
Can by the collection of illustrative plates contrasted under 250nm, 260nm, 270nm, 300nm, 280nm, 290nm, 325nm, 360nm wavelength
To find, each peak appearance situation of chromatogram under 270nm wavelength is more balanced, and lobetyolin's maximum absorption wavelength is in 270nm.Therefore
Selection 270nm leads to Detection wavelength during particle fingerprint map analyzing as train of thought.
Methodological study
(1) precision is investigated
Take the train of thought that lot number is 16102381 to lead to particle 2.0g, need testing solution is prepared according to test sample preparation method, even
Continue sample introduction 6 times, record chromatogram, and similarity-rough sets are carried out to 6 collection of illustrative plates.As a result the similarity between collection of illustrative plates is 1.000, table
Bright instrument precision is good, is shown in Table 2.
The train of thought of table 2 leads to particle Precision Experiment result (16102381)
(2) repeatability is investigated
Take the train of thought that lot number is 16102381 to lead to particle, 6 parts of need testing solutions are prepared according to test sample preparation method.According to
Above-mentioned chromatographic condition sample introduction, chromatogram is recorded, and similarity-rough set is carried out to 6 collection of illustrative plates.As a result the similarity between collection of illustrative plates is
1.000, show that this method repeatability is good, be shown in Table 3.
The train of thought of table 3 leads to particle repeated experiment result (16102381)
(3) study on the stability
Take the train of thought that lot number is 16102381 to lead to particle 2.0g, need testing solution is prepared according to test sample preparation method, point
0h, 2h, 4h, 8h, 12h, 16h, 20h, 24h sample introduction not after sample preparation, chromatogram is recorded, and phase is carried out to the full peak of 8 collection of illustrative plates
Compare like degree.As a result the similarity between collection of illustrative plates is 1.000, shows that sample stability is good, is shown in Table 4.
The train of thought of table 4 leads to granule stability experimental result (16102381)
Similarity-rough set
The HPLC collection of illustrative plates that collect 14 batches of train of thoughts are led to particle is imported successively with AIA forms《Chromatographic fingerprints of Chinese materia medica
Similarity evaluation system》, similarity comparison is carried out with the reference fingerprint of generation, the results are shown in Table 5.
5 14 batches of train of thoughts of table lead to particulate samples similarity evaluation result
The similarity of above-mentioned as shown by data, the logical particle of 14 batches of train of thoughts and reference fingerprint illustrates product more than 0.9
Quality stability it is good.
Compared with prior art, train of thought provided by the invention leads to particle fingerprint atlas detection method stability and reproducible,
The quality of particle can be led to control train of thought and ensure clinical efficacy with quality that is objective, comprehensively and accurately evaluating the logical particle of train of thought
It is significant.
Described above is only exemplary embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member for, under the premise without departing from the principles of the invention, can also be made according to prior art improvement and optimization, these improve and
Optimization falls within protection scope of the present invention.
Claims (7)
1. a kind of train of thought leads to the fingerprint atlas detection method of particle, including leads to particle progress to train of thought using high performance liquid chromatography
Detection, wherein, chromatographic condition includes：
Using octadecylsilane chemically bonded silica as filler, using acetonitrile as mobile phase A, using 0.3 volume % aqueous formic acids as stream
Dynamic phase B, gradient elution, 0.5~1.5mlmin of flow velocity-1, 20~35 DEG C of column temperature, 210~360nm of Detection wavelength；Theoretical tray
Number is calculated by rutin peak should be not less than 3000.
2. detection method as claimed in claim 1, wherein, during gradient elution, the ratio of mobile phase A, which becomes, to be turned to：0~
3min, 5 volume % acetonitriles；The volume % acetonitriles of 3~50min, 5 volume %~42；The body of 50~50.1min, 42 volume %~95
Product % acetonitriles；50.1~60min, 95 volume % acetonitriles；That is gradient elution program such as following table：
3. detection method as claimed in claim 1, wherein, the chromatographic column using octadecylsilane chemically bonded silica as filler is
Phenomenenx luna (2) C18,4.6*250mm, 5um.
It is 4. further comprising the steps of such as the detection method any one of claim 1-3：
(1) preparation of need testing solution：Take train of thought to lead to particle 2.0g, it is accurately weighed, put in 100ml conical flask with cover, precision plus
Enter 70 volume 25~75ml of % ethanol, close plug, weighed weight, be ultrasonically treated 10~40 minutes under conditions of 250w, 40KHz,
Let cool, the weight of less loss is supplied with 70 volume % ethanol, is shaken up, filter, take subsequent filtrate, produce；
(2) preparation of reference substance solution：Puerarin, rutin, forulic acid, lobetyolin, tanshin polyphenolic acid B, Quercetin, Kaempferide is taken to fit
Amount, add 70 volume % ethanol that every 1ml is made containing μ g of Puerarin 112, μ g of rutin 204, μ g of forulic acid 44, μ g of lobetyolin 32, pellet
μ g of phenolic acid B 92, μ g of Quercetin 40, the μ g of Kaempferide 28 mixed reference substance solution, are produced；
(3) determine：It is accurate respectively to draw reference substance solution and each 2~10 μ l of need testing solution, inject high performance liquid chromatograph, note
The chromatogram in 60 minutes is recorded, is used《Similarity evaluation》2004A versions are handled collection of illustrative plates, i.e.,
Obtain the finger-print that train of thought leads to particle.
5. a kind of train of thought leads to the fingerprint atlas detection method of particle, comprise the following steps：
(1) preparation of need testing solution：Take train of thought to lead to particle 2.0g, it is accurately weighed, put in 100ml conical flask with cover, precision plus
Enter 70 volume 25~75ml of % ethanol, close plug, weighed weight, be ultrasonically treated 10~40 minutes under conditions of 250w, 40KHz,
Let cool, the weight of less loss is supplied with 70 volume % ethanol, is shaken up, filter, take subsequent filtrate, produce；
(2) preparation of reference substance solution：Puerarin, rutin, forulic acid, lobetyolin, tanshin polyphenolic acid B, Quercetin, Kaempferide is taken to fit
Amount, add 70 volume % ethanol that every 1ml is made containing μ g of Puerarin 112, μ g of rutin 204, μ g of forulic acid 44, μ g of lobetyolin 32, pellet
μ g of phenolic acid B 92, μ g of Quercetin 40, the μ g of Kaempferide 28 mixed reference substance solution, are produced；
(3) determine：It is accurate respectively to draw reference substance solution and each 2~10 μ l of need testing solution, inject high performance liquid chromatograph, note
The chromatogram in 60 minutes is recorded, is used《Similarity evaluation》2004A versions are handled collection of illustrative plates, i.e.,
Obtain the finger-print that train of thought leads to particle；
Wherein, in step (3), the chromatographic condition of high performance liquid chromatography measure is：Using octadecylsilane chemically bonded silica as filling
Agent, using acetonitrile as mobile phase A, using 0.3 volume % aqueous formic acids as Mobile phase B, gradient elution, 0.5~1.5ml of flow velocity
min-1, 20~35 DEG C of column temperature, 210~360nm of Detection wavelength；Theoretical cam curve is calculated by rutin peak should be not less than 3000.
6. the train of thought obtained by the fingerprint atlas detection method of the logical particle of train of thought as any one of claim 1-5 is led to
Particle finger-print, there are 21 shared peaks in the collection of illustrative plates.
7. finger-print as claimed in claim 6, wherein, the retention time at each shared peak is respectively：Shared peak 1：
13.234min；Shared peak 2：15.952min；Shared peak 3：17.602min；Shared peak 4：20.612min；Shared peak 5：
21.099min；Shared peak 6：21.321min；Shared peak 7：24.057min；Shared peak 8：27.190min；Shared peak 9：
28.845min；Shared peak 10：29.452min；Shared peak 11：29.830min；Shared peak 12：30.206min；Shared peak 13：
33.715min；Shared peak 14：34.979min；Shared peak 15：37.136min；Shared peak 16：38.789min；Shared peak 17：
39.278min；Shared peak 18：39.610min；Shared peak 19：41.671min；Shared peak 20：45.049min；Shared peak 21：
48.173min。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710733401.1A CN107589185B (en) | 2017-08-24 | 2017-08-24 | Fingerprint detection method of Mailuotong granules and fingerprint thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710733401.1A CN107589185B (en) | 2017-08-24 | 2017-08-24 | Fingerprint detection method of Mailuotong granules and fingerprint thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108845059A (en) * | 2018-08-24 | 2018-11-20 | 鲁南制药集团股份有限公司 | The method for building up and its standard diagram of train of thought Shutong granules HPLC finger-print |
| CN109406672A (en) * | 2018-11-20 | 2019-03-01 | 合肥创新医药技术有限公司 | A kind of fingerprint spectrum method using HPLC measurement Chinese medicine yiguan decoction |
| CN112748201A (en) * | 2020-12-28 | 2021-05-04 | 南京海昌中药集团有限公司 | Method for detecting fingerprint of radix puerariae assorted Chinese herbal tea |
| CN113720954A (en) * | 2021-09-18 | 2021-11-30 | 安徽中医药大学第一附属医院(安徽省中医院) | Method for establishing fingerprint spectrum of brain-improving capsule and quality evaluation method |
| CN113759009A (en) * | 2020-08-26 | 2021-12-07 | 北京康仁堂药业有限公司 | Method for constructing characteristic spectrum of equisetum hiemale and preparation thereof |
| CN114166958A (en) * | 2021-10-29 | 2022-03-11 | 合肥创新医药技术有限公司 | Fingerprint detection method and application of traditional Chinese medicine compound cang huo pingwei granules |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1817365A (en) * | 2005-10-31 | 2006-08-16 | 石家庄制药集团欧意药业有限公司 | Injecting Mailuoning, its preparation and quality control thereof |
| CN101744920A (en) * | 2010-02-04 | 2010-06-23 | 国药控股深圳中药有限公司 | Fingerprint control method for Jian'er Qinjie oral solution |
| CN102068657A (en) * | 2009-11-20 | 2011-05-25 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Quality control method of venation relaxing pill as Chinese herbal preparation |
| CN103197004A (en) * | 2012-12-24 | 2013-07-10 | 扬子江药业集团南京海陵药业有限公司 | Quality control method for Mailuotong capsule(capsule dredging main and collateral channels) |
-
2017
- 2017-08-24 CN CN201710733401.1A patent/CN107589185B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1817365A (en) * | 2005-10-31 | 2006-08-16 | 石家庄制药集团欧意药业有限公司 | Injecting Mailuoning, its preparation and quality control thereof |
| CN102068657A (en) * | 2009-11-20 | 2011-05-25 | 天津中新药业集团股份有限公司乐仁堂制药厂 | Quality control method of venation relaxing pill as Chinese herbal preparation |
| CN101744920A (en) * | 2010-02-04 | 2010-06-23 | 国药控股深圳中药有限公司 | Fingerprint control method for Jian'er Qinjie oral solution |
| CN103197004A (en) * | 2012-12-24 | 2013-07-10 | 扬子江药业集团南京海陵药业有限公司 | Quality control method for Mailuotong capsule(capsule dredging main and collateral channels) |
Non-Patent Citations (2)
| Title |
|---|
| 卓菊等: "《中药制剂检测技术》", 31 January 2013, 中国医药科技出版社 * |
| 谭琴等: "脉络通胶囊质量标准", 《中国实验方剂学杂志》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108845059A (en) * | 2018-08-24 | 2018-11-20 | 鲁南制药集团股份有限公司 | The method for building up and its standard diagram of train of thought Shutong granules HPLC finger-print |
| CN108845059B (en) * | 2018-08-24 | 2020-05-26 | 鲁南制药集团股份有限公司 | Method for establishing HPLC fingerprint of Mailuoshutong granules and standard spectrum thereof |
| CN109406672A (en) * | 2018-11-20 | 2019-03-01 | 合肥创新医药技术有限公司 | A kind of fingerprint spectrum method using HPLC measurement Chinese medicine yiguan decoction |
| CN113759009A (en) * | 2020-08-26 | 2021-12-07 | 北京康仁堂药业有限公司 | Method for constructing characteristic spectrum of equisetum hiemale and preparation thereof |
| CN113759009B (en) * | 2020-08-26 | 2023-03-17 | 北京康仁堂药业有限公司 | Method for constructing characteristic spectrum of equisetum hiemale and preparation thereof |
| CN112748201A (en) * | 2020-12-28 | 2021-05-04 | 南京海昌中药集团有限公司 | Method for detecting fingerprint of radix puerariae assorted Chinese herbal tea |
| CN113720954A (en) * | 2021-09-18 | 2021-11-30 | 安徽中医药大学第一附属医院(安徽省中医院) | Method for establishing fingerprint spectrum of brain-improving capsule and quality evaluation method |
| CN113720954B (en) * | 2021-09-18 | 2022-02-11 | 安徽中医药大学第一附属医院(安徽省中医院) | Method for establishing fingerprint spectrum of brain-improving capsule and quality evaluation method |
| CN114166958A (en) * | 2021-10-29 | 2022-03-11 | 合肥创新医药技术有限公司 | Fingerprint detection method and application of traditional Chinese medicine compound cang huo pingwei granules |
| CN114166958B (en) * | 2021-10-29 | 2024-03-29 | 合肥创新医药技术有限公司 | Detection method of fingerprint of traditional Chinese medicine compound herba xanthil stomach-calming particles and application thereof |
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