CN104897831B - The construction method of NAOXINTONG finger printing - Google Patents
The construction method of NAOXINTONG finger printing Download PDFInfo
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Abstract
The embodiment of the invention discloses the construction method of a kind of NAOXINTONG finger printing, comprise the following steps: (1), prepare need testing solution；(2), Ultra Performance Liquid Chromatography detection；(3), NAOXINTONG finger printing is built.Construction method precision height, stability and the favorable reproducibility of the NAOXINTONG finger printing that the present invention obtains, method simplicity, can evaluate the quality of NAOXINTONG quickly and accurately.
Description
Technical field
The present invention relates to the field of quality control of Chinese medicine preparation, particularly to the structure side of NAOXINTONG finger printing
Method.
Background technology
Chinese medicine fingerprint refer to some Chinese crude drug or Chinese medicine preparation appropriately processed after, use certain analysis
Means, the chromatogram that can indicate its chemical feature obtained or spectrogram.Chinese medicine fingerprint quality control
Technology, with chromatography as means, by the com-parison and analysis to fingerprint characteristic, obtains Chinese crude drug and Chinese medicine
The relevant information of intermediate formulations composition, evaluates the verity of its quality, Optimality and stability, is existing
The best techniques of stage overall merit Chinese medicine quality.State Food and Drug Administration has required Chinese medicine preparation
Implement finger printing quality control.
" country's standard for traditional Chinese medicines compilation-Chinese patent medicine place mark of National Drug Administration's compilation in 2002
Accurate rise national standard part " " internal medicine-brain system " fascicle, disclose the quality standard of NAOXINTONG, (mark
Accurate number: WS-10001 (ZD-0001)-2002) mainly contain 16 taste medical materials, the parts by weight between each medical material
Ratio is: the Radix Astragali 66 parts, Radix Paeoniae Rubra 27 parts, Radix Salviae Miltiorrhizae 27 parts, Radix Angelicae Sinensis 27 parts, Rhizoma Chuanxiong 27 parts, 27 parts of Semen Persicae,
13 parts of Flos Carthami, Olibanum (system) 13 parts, Myrrha (processed) 13 parts, Caulis Spatholobi 20 parts, Radix Achyranthis Bidentatae 27 parts, Ramulus Cinnamomi 20
Part, Ramulus Mori 27 parts, Pheretima 27 parts, Scorpio 13 parts, Hirudo 27 parts.Have expectorant repercussive, blood vessels and
Freely, numbness preferably pain only effect, be used for treating coronary heart diseases and angina pectoris.At present for this medicine quality control side
Method is: use Ultra Performance Liquid Chromatography (UPLC) technology to measure Hydroxy Carthamus yellow in NAOXINTONG JIAONANG simultaneously
A, peoniflorin, ferulic acid, salvianolic acid B, 5 kinds of main chemical compositions of ligustilide.But, in NAOXINTONG
There is also other effective ingredient substantial amounts of, and these effective ingredient are also to have weight for the quality control of NAOXINTONG
Want meaning.Accordingly, it would be desirable to rebuild NAOXINTONG finger printing, to reflect contained by NAOXINTONG all sidedly
Chemical effective ingredient, preferably control the medicine quality of NAOXINTONG.
Summary of the invention
For solving the problems referred to above, the embodiment of the invention discloses the construction method of a kind of NAOXINTONG finger printing.
Technical scheme is as follows:
1, the construction method of a kind of NAOXINTONG finger printing, comprises the following steps:
(1), need testing solution is prepared
Take NAOXINTONG powder, extract with volume fraction 10-90% methanol solution, constant volume, obtain need testing solution；
(2), Ultra Performance Liquid Chromatography detection
Need testing solution is injected in Ultra Performance Liquid Chromatography instrument, detect, obtain NAOXINTONG test sample
Chromatogram；Wherein chromatographic test strip part includes: with octadecyl silane post for fixing phase, with 100% second
Nitrile: 100% methanol volume ratio be the acetonitrile methanol mixed solution of 1: 1 be A phase, volume fraction 0.1% phosphate aqueous solution
For B phase, composition flowing phase；Use gradient elution；Detection wavelength: 210-400nm；Flow velocity: 0.1-0.5mL/min；
Column temperature: 25-40 DEG C；Sample size: 1-5 μ L；
(3), NAOXINTONG finger printing is built
The chromatogram of NAOXINTONG test sample is imported " similarity evaluation " carry out
Analyze, obtain NAOXINTONG finger printing.
In the preferred embodiment of the present invention, described prepare need testing solution step be: weighed matter
Amount is the NAOXINTONG powder of M, adds the volume fraction 10-90% methanol solution of V volume, obtains the two mixed liquor,
Weight M of weighed mixed liquor1, extract, be re-weighed, supply with respective concentration methanol weightless to M1, it is centrifugal,
Take supernatant, obtain need testing solution.
In the preferred embodiment of the present invention, the volume fraction of described methanol solution is 75%.
In the preferred embodiment of the present invention, it is extracted as supersound extraction, supersound extraction 35min described in.
In the preferred embodiment of the present invention, described NAOXINTONG powder quality and described volume fraction 75%
The volume ratio of methanol solution is 1: 40.
In the one more preferably embodiment of the present invention, by described need testing solution with ultra-pure water with volume
Ratio is 5: 2 dilutions, then carries out Ultra Performance Liquid Chromatography detection.
In the preferred embodiment of the present invention, described elution program: 0-2min, A phase: B phase
=87: 13；2-7min, A phase: B phase=(87-78): (13-22)；7-25min, A phase: B phase=(78-60): (22-40)；
25-27min, A phase: B phase=(60-45): (40-55)；27-30min, A phase: B phase=45: 55；30-39min,
A phase: B phase=(45-10): (55-90)；39-40min, A phase: B phase=(10-87): (90-13)；Described column temperature is
40℃；Described flow velocity: 0.2mL/min；Described sample size: 2 μ L.
In the one more preferably embodiment of the present invention, described detection wavelength is 324nm.
In the preferred embodiment of the present invention, described NAOXINTONG finger printing has 24 fingerprint peakses,
Relative retention time is respectively as follows: No. 1 peak: 0.098-0.100；No. 2 peak: 0.202-0.202；No. 3 peaks:
0.238-0.257；No. 4 peaks: relative retention time 0.330-0.347；No. 5 peak: 0.481-0.497；No. 6 peaks:
0.523-0.538；No. 7 peak: 0.619-0.639；No. 8 peak: 0.648-0.669；No. 9 peak: 0.710-0.731；
No. 10 peak: 0.724-0.745；No. 11 peak: 0.752-0.773；No. 12 peak: 0.784-0.807；No. 13 peaks:
0.804-0.828；No. 14 peak: 0.857-0.880；No. 15 peak: 0.895-0.919；No. 16 peaks are with reference to peak: 1；
No. 17 peak: 1.087-1.110；No. 18 peak: 1.620-1.632；No. 19 peak: 1.726-1.737；No. 20 peaks:
1.744-1.755；No. 21 peak: 1.897-1.904.；No. 22 peak: 1.909-1.916.；No. 23 peak: 1.941-1.948；
No. 24 peak: 1.990-1.997.
The construction method precision of the NAOXINTONG finger printing that the present invention obtains is high, stability and favorable reproducibility,
Method is easy, can evaluate the quality of NAOXINTONG quickly and accurately.
Accompanying drawing explanation
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to enforcement
In example or description of the prior art, the required accompanying drawing used is briefly described, it should be apparent that, describe below
In accompanying drawing be only some embodiments of the present invention, for those of ordinary skill in the art, do not paying
On the premise of going out creative work, it is also possible to obtain other accompanying drawing according to these accompanying drawings.
Fig. 1 is the finger printing of the NAOXINTONG of 28 batches；
Fig. 2 is the NAOXINTONG reference fingerprint of the NAOXINTONG collection of illustrative plates matching of 28 batches.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, the technical scheme in the embodiment of the present invention is carried out clearly
Chu, be fully described by, it is clear that described embodiment be only a part of embodiment of the present invention rather than
Whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not making creation
The every other embodiment obtained under property work premise, broadly falls into the scope of protection of the invention.
Technical scheme, for preparing need testing solution, selection volume fraction 10-90% methanol is molten
Liquid extracts the chemical composition in NAOXINTONG powder, it is achieved the mode fully extracted and time, the technology of this area
Personnel can select according to practical experience and determine.After extraction, constant volume is the conventional means of this area.Preferably,
After NAOXINTONG powder and volume fraction 10-90% methanol mixed, weigh, after extraction, be re-weighed, use
The methanol of same volume mark supplies weightlessness, then takes supernatant, can reach two purposes: fully extract and constant volume.
This constant volume method, relative to exhausting supernatant again by the method for Extraction solvent constant volume, can eliminate experimental implementation process
In supernatant artificially all can not be exhausted produced error, it is thus achieved that result closer to theoretical actual value.
This constant volume method is the improvement to traditional constant volume method.After the method constant volume, take part supernatant, energy
Enough meet the demand of subsequent experimental.Simplify the experimental procedure of constant volume.Certainly, above-mentioned preparation NAOXINTONG
The method of need testing solution is not the method uniquely preparing need testing solution, and those skilled in the art completely may be used
With according to practice, use other methods preparing NAOXINTONG need testing solution.For extracting NAOXINTONG powder
The reagent at end, those skilled in the art can select in ethanol, methanol, n-butyl alcohol equal solvent according to practical experience
One or more.
In the inventive solutions, it is volume fraction prepare need testing solution step preferably extracting reagent
75% methanol, supersound extraction, supersound extraction time 35min, NAOXINTONG quality is molten with volume fraction 75% methanol
The volume ratio of liquid is 1: 40 (mg/mL), optimizes the technique that methanol solution extracts NAOXINTONG, it is achieved efficiently, soon
The purpose of chemical composition in speed, fully extraction NAOXINTONG.
Need testing solution and ultra-pure water are preferably 5: 2 dilutions with volume ratio by the present invention, then carry out ultra high efficiency liquid phase
Chromatograph detects, it is therefore intended that: change the polarity of solvent, reduce solvent and flowing phase polarity difference, thus change
Kind peak shape, it is thus achieved that high-quality chromatogram.
In the inventive solutions, Ultra Performance Liquid Chromatography testing conditions includes: be bonded with octadecyl
Silicagel column is fixing phase, with 100% acetonitrile: 100% methanol volume ratio is that the acetonitrile methanol mixed solution of 1: 1 is as A
Phase, volume fraction 0.1% phosphate aqueous solution is B phase, composition flowing phase；Use gradient elution；Detection wavelength:
210-400nm；Flow velocity: 0.1-0.5mL/min；Column temperature: 25-40 DEG C；Sample size: 1-5 μ l.Wherein eluting
Program: 0-2min, A phase: B phase=87: 13；2-7min, A phase: B phase=(87-78): (13-22)；7-25min, A
Phase: B phase=(78-60): (22-40)；25-27min, A phase: B phase=(60-45): (40-55)；27-30min, A phase: B
Phase=45: 55；30-39min, A phase: B phase=(45-10): (55-90)；39-40min, A phase: B phase
=(10-87): (90-13).Above-mentioned flowing phase is selected to be advantageous in that: in need testing solution, each composition all obtains well
Peak shape and separating degree；Above-mentioned elution program is used to be advantageous in that: each composition separating degree in need testing solution
Good, the detection time is short, highly sensitive.
In technical scheme, the wave-length coverage of chromatograph detection is at 210-400nm, and this wave-length coverage covers
The absorbing wavelength of all of chemical composition in NAOXINTONG medicine, preferably detects under 324nm wavelength, is advantageous in that:
At that wavelength, each composition in need testing solution all has good response.
The chromatogram of NAOXINTONG test sample is imported " similarity evaluation " carry out
Analyze, obtain NAOXINTONG finger printing.
Term " accurately weighed " mentioned by the present invention: accurately weighed mean weigh weight should be accurately to be taken heavily
The one thousandth of amount." precision measures ": refer to that the accuracy measuring volume should meet in national standard this metering
The precise requirements of utensil.
10-90% methanol solution mentioned by the present invention, 75% methanol, 0.1% phosphate aqueous solution are all volume integral
Number.
In the art, the relative standard deviation (RSD) of the experimental data of assay, within 5%, is said
Bright experimental result is true and reliable.
Instrument and reagent
U.S.'s WATERS Ultra Performance Liquid Chromatography (UPLC) ACCURICY system, including quaternary supertension
Solvent system, auto injection constant temperature sample management device, diode array (PDA) detector, Empower 3
Chromatographic work station；Millipore company Mill-Q II type Superpure water machine；Sigma Co., USA 3K15 is at a high speed
Centrifuge；Mettler Toledo company of Switzerland AX205 100,000/balance；Ningbo new sesame biotechnology has
Limit company SB-1000YDTD ultrasonic cleaner.
Methanol, acetonitrile (Fisher company of the U.S.), ultra-pure water (Millipore ultra-pure water cleaning system prepares),
Phosphoric acid (chromatographic grade).
Reference substance chlorogenic acid, S-A Hydroxysafflor yellow A, ferulic acid, five galloyl glucoses, 3,5-bis-
Caffeoylquinic acids, 1,5-Dicaffeoylquinic acid, FNS, rosmarinic acid, alkannic acid,
Salvianolic acid B, calycosin, ligustilide, butylidene phthalide, cryptotanshinone are purchased from Chengdu, Sichuan graceful
Si Te Bioisystech Co., Ltd.
NAOXINTONG JIAONANG (totally 28 batches, lot number 120756,120757,120759,120760,120761,
120763、120764、120767、120770、120771、120773、120774、120775、120777、
120778、120779、120781、120783、120784、120785、120787、120788、120801、
120803,120804,120805,120819,120820) (0.4g/ is provided by Shaanxi Buchang Pharmaceuticals Co., Ltd.
Grain).
Embodiment 1
(1), the preparation of need testing solution
The NAOXINTONG powder 0.25g of accurately weighed 28 batches, adds 75% methanol, is settled to respectively respectively
10mL, weighs NAOXINTONG powder and the weight of 75% methyl alcohol mixed liquor, supersound extraction 35min respectively, put to
Room temperature, then weigh the weight of mixed liquor, supplying weightlessness with 75% methanol, shake up, centrifugal 14000rpm is centrifuged
10min, centrifugal 2 times, takes supernatant, obtains the NAOXINTONG need testing solution of 28 batches.
(2), Ultra Performance Liquid Chromatography detection
It is 5: 2 dilutions with ultra-pure water with volume ratio respectively by the NAOXINTONG need testing solution of 28 batches, then enters
Row UPLC detects, and chromatographic condition includes: with octadecyl silane post for fixing phase, with 100% second
Nitrile: 100% methanol volume ratio be the acetonitrile methanol mixed solution of 1: 1 be A phase, 0.1% phosphate aqueous solution is B phase,
Composition flowing phase, 0-2min, A phase: B phase=87: 13；2-7min, A phase: B phase=(87-78): (13-22)；
7-25min, A phase: B phase=(78-60): (22-40)；25-27min, A phase: B phase=(60-45): (40-55)；
27-30min, A phase: B phase=45: 55；30-39min, A phase: B phase=(45-10): (55-90)；39-40min,
A phase: B phase=(10-87): (90-13)；Detection wavelength is 324nm；Flow velocity 0.2mL/min；Column temperature 40 DEG C；
Sample size 2 μ L.Theoretical cam curve salvianolic acid B (324nm)=5.54 (tR/Wh/2)2=324850.3.Obtain 28
The NAOXINTONG chromatogram of batch.28 batch NAOXINTONG retention times and peak area are shown in Table 1.
1 28 batch NAOXINTONG retention times of table and peak area area Results
Relative retention time and relative Retention area with No. 16 salvianolic acid B characteristic peaks, for 1, calculate other peaks
Relative retention time and relative peak area, the results are shown in Table 2.
The relative retention time at table 2 characteristic fingerprint peak and relative peak area
(3), the structure of NAOXINTONG finger printing
The NAOXINTONG chromatogram of 28 batches is imported " similarity evaluation "
(2004A) version is analyzed, and obtains the NAOXINTONG finger printing (as shown in Figure 1) and 28 of 28 batches
The NAOXINTONG reference fingerprint (as shown in Figure 2) of the NAOXINTONG chromatograph matching of individual batch, NAOXINTONG feature refers to
Stricture of vagina peak has 24, and its peak area accounts for more than the 80% of total peak area.NAOXINTONG color with 120756 batches
Spectrogram is reference, and the NAOXINTONG of other batches compares therewith, and similarity is all higher than 0.971；Comparison with matching
Finger printing is reference, and the NAOXINTONG of 28 batches compares therewith, and similarity is all higher than 0.976 (such as table 3 institute
Show), the construction method of NAOXINTONG finger printing of the present invention is described accurately and reliably.
Table 3 similarity result
Hereinafter choose the NAOXINTONG JIAONANG that batch is 120805, by precision, repeatability and stability test,
Technical scheme is carried out Method validation.
(4) preparation of reference substance solution, is mixed
Accurately weighed S-A Hydroxysafflor yellow A (HSYA), five galloyl glucoses, kaempferol respectively
-3-O-rutinoside, alkannic acid, salvianolic acid B, calycosin, ligustilide, butylidene phthalide compare
Product 2mg, adds 100% methanol dissolving and is settled to 1mL, be configured to the various reference substances that concentration is 2mg/mL female
Liquid；Respectively accurately weighed chlorogenic acid, ferulic acid, 3,5-diCQA, 1,5-Dicaffeoylquinic acid,
Rosmarinic acid, cryptotanshinone reference substance 1mg, add 100% methanol dissolving and be settled to 1mL, and being configured to concentration is
The various reference substance mother solutions of 1mg/mL.
Respectively precision measure chlorogenic acid, S-A Hydroxysafflor yellow A, ferulic acid, five galloyl glucoses,
3,5-diCQA, 1,5-Dicaffeoylquinic acid, FNS, rosmarinic acid,
Alkannic acid, salvianolic acid B, calycosin, ligustilide, butylidene phthalide and cryptotanshinone reference substance mother solution
60μL、60μL、30μL、60μL、30μL、45μL、45μL、60μL、120μL、90μL、50μL、
60 μ L, 45 μ L and 60 μ L, 100% methanol constant volume to 1mL.Reference substance mother solution must be mixed.Its medium green is former
Acid concentration is 60 μ g/mL, S-A Hydroxysafflor yellow A concentration is 120 μ g/mL, ferulic acid concentration is
30 μ g/mL, five galloyl glucose concentration are 120 μ g/mL, 3,5-diCQA concentration is
30 μ g/mL, 1,5-Dicaffeoylquinic acid concentration are 45 μ g/mL, FNS concentration is
90 μ g/mL, rosmarinic acid concentration are 60 μ g/mL, alkannic acid concentration is 240 μ g/mL, salvianolic acid B concentration
Be 180 μ g/mL, calycosin concentration be 100 μ g/mL, ligustilide concentration be 120 μ g/mL, butylene
Base Phthalide concentration is 90 μ g/mL, cryptotanshinone concentration is 60 μ g/mL, by mother solution according to 1/2,1/5,1/10,
1/25,1/50,1/125,1/250,1/300 times, with 100% methanol stepwise dilution, obtain respective concentration
Mixing reference substance solution (being shown in Table 4), is injected separately into chromatograph of liquid, and sample introduction 2 μ L is measured, and draws
To standard curve, the results are shown in Table 5.
Table 4 mixes the concentration (μ g/mL) of reference substance solution
Table 5 each reference substance standard curve
(5), precision test
Accurately weighed batch is NAOXINTONG JIAONANG 0.25g of 120805, extracts to enter by embodiment 1 step (1)
OK, chromatograph detection is carried out by embodiment 1 step (2).Sample introduction 2 μ L is measured.Calculate chlorogenic acid, hydroxyl
Base safflower yellow A, ferulic acid, five galloyl glucoses, 3,5-diCQA, 1,5-bis-
Caffeoylquinic acids, FNS, rosmarinic acid, alkannic acid, salvianolic acid B, Mao Ruiyi
Flavone, ligustilide, butylidene phthalide, the content of cryptotanshinone, RSD is respectively less than 5%, meets precision
Requirement, the results are shown in Table 6.
(6), stability test
Accurately weighed batch is NAOXINTONG JIAONANG 0.25g of 120805, extracts to enter by embodiment 1 step (1)
OK, chromatograph detection is carried out by embodiment 1 step (2).Respectively at 0h, 1h, 2h, 4h, 6h, 8h, 10h,
12h, 24h, be injected separately into chromatograph of liquid, and sample introduction 2 μ L is measured, and investigates stablizing of need testing solution
Property.Calculate chlorogenic acid, S-A Hydroxysafflor yellow A, ferulic acid, five galloyl glucoses, 3,5-bis-coffee
Coffee acyl quininic acid, 1,5-Dicaffeoylquinic acid, FNS, rosmarinic acid, alkannic acid,
Salvianolic acid B, calycosin, ligustilide, butylidene phthalide, the content of cryptotanshinone, RSD is the least
In 5%.It is basicly stable that result shows that need testing solution room temperature places 24h, the results are shown in Table 7.
(7), replica test
Accurately weighed batch is NAOXINTONG JIAONANG 0.25g of 120805, extracts to enter by embodiment 1 step (1)
OK, 6 parts of samples of parallel processing, chromatograph detection is carried out by embodiment 1 step (2).It is injected separately into liquid phase color
Spectrometer, sample introduction 2 μ L is measured, and investigates the repeatability of experiment.Calculate chlorogenic acid, Hydroxy Carthamus yellow
A, ferulic acid, five galloyl glucoses, 3,5-diCQA, 1,5-Dicaffeoylquinic acid,
FNS, rosmarinic acid, alkannic acid, salvianolic acid B, calycosin, ligustilide,
Butylidene phthalide, the content of cryptotanshinone, RSD is respectively less than 5%.Result shows the construction method weight of the present invention
Renaturation is good, the results are shown in Table 8.
Above-mentioned precision, repeatability and stability test result show, the construction method of NAOXINTONG finger printing
There is precision height, stability and favorable reproducibility, the feature of method simplicity, brain can be differentiated quickly and accurately
The quality that the heart is logical.
In the art, Chinese medicine chromatogram is compared with reference fingerprint, and similarity is more than 0.9, i.e. can determine that
The construction method of this finger printing is accurately and reliably.Accordingly, for the quality testing of NAOXINTONG, can be with 28
The reference fingerprint of the NAOXINTONG chromatograph matching of batch is standard, evaluates the quality of NAOXINTONG.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit protection scope of the present invention.
All any modification, equivalent substitution and improvement etc. made within the spirit and principles in the present invention, are all contained in
In protection scope of the present invention.
Claims (9)
1. the construction method of a NAOXINTONG finger printing, it is characterised in that comprise the following steps:
(1), need testing solution is prepared
Take NAOXINTONG powder, extract with volume fraction 10-90% methanol solution, constant volume, obtain need testing solution；
(2), Ultra Performance Liquid Chromatography detection
Need testing solution is injected in Ultra Performance Liquid Chromatography instrument, detect, obtain NAOXINTONG test sample
Chromatogram；Wherein chromatographic test strip part includes: with octadecyl silane post for fixing phase, with 100% second
Nitrile: 100% methanol volume ratio be the acetonitrile methanol mixed solution of 1:1 be A phase, volume fraction 0.1% phosphate aqueous solution
For B phase, composition flowing phase；Use gradient elution；Detection wavelength: 210-400nm；Flow velocity: 0.1-0.5mL/min；
Column temperature: 25-40 DEG C；Sample size: 1-5 μ L；Wherein elution program: 0-2min, A phase: B phase=87:13；2-7min,
A phase: B phase=(87-78): (13-22)；7-25min, A phase: B phase=(78-60): (22-40)；25-27min, A phase: B
Phase=(60-45): (40-55)；27-30min, A phase: B phase=45:55；30-39min, A phase: B phase
=(45-10): (55-90)；39-40min, A phase: B phase=(10-87): (90-13)；
(3), NAOXINTONG finger printing is built
The chromatogram of NAOXINTONG test sample is imported " similarity evaluation " carry out
Analyze, obtain NAOXINTONG finger printing.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 1, it is characterised in that institute
State and prepare the step of need testing solution and be: weighed quality is the NAOXINTONG powder of M, adds the volume integral of V volume
Number 10-90% methanol solution, obtains the two mixed liquor, weight M of weighed mixed liquor1, extract, be re-weighed,
Supply weightless to M with respective concentration methanol1, centrifugal, take supernatant, obtain need testing solution.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 2, it is characterised in that institute
The volume fraction of the methanol solution stated is 75%.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 2, it is characterised in that institute
State and be extracted as supersound extraction, supersound extraction 35min.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 3, it is characterised in that institute
The volume ratio stating NAOXINTONG powder quality and volume fraction 75% methanol solution is 1mg:40mL.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 5, it is characterised in that will
Described need testing solution dilutes for 5:2 with volume ratio with ultra-pure water, then carries out Ultra Performance Liquid Chromatography detection.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 1, it is characterised in that institute
Stating column temperature is 40 DEG C；Described flow velocity: 0.2mL/min；Described sample size: 2 μ L.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 5, it is characterised in that institute
Stating detection wavelength is 324nm.
The construction method of a kind of NAOXINTONG finger printing the most as claimed in claim 1, it is characterised in that institute
Stating NAOXINTONG finger printing and have 24 fingerprint peakses, relative retention time is respectively as follows: No. 1 peak: 0.098-0.100；
No. 2 peak: 0.202-0.202；No. 3 peak: 0.238-0.257；No. 4 peaks: relative retention time 0.330-0.347；
No. 5 peak: 0.481-0.497；No. 6 peak: 0.523-0.538；No. 7 peak: 0.619-0.639；No. 8 peak: 0.648-0.669；
No. 9 peak: 0.710-0.731；No. 10 peak: 0.724-0.745；No. 11 peak: 0.752-0.773；No. 12 peaks:
0.784-0.807；No. 13 peak: 0.804-0.828；No. 14 peak: 0.857-0.880；No. 15 peak: 0.895-0.919；
No. 16 peaks are with reference to peak: 1；No. 17 peak: 1.087-1.110；No. 18 peak: 1.620-1.632；No. 19 peaks:
1.726-1.737；No. 20 peak: 1.744-1.755；No. 21 peak: 1.897-1.904.；No. 22 peak: 1.909-1.916.；
No. 23 peak: 1.941-1.948；No. 24 peak: 1.990-1.997.
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| CN107422052B (en) * | 2017-06-29 | 2020-06-05 | 天津中医药大学 | Green extraction and determination method for various compounds in Chinese herbal compound Naoxintong |
| CN109307721B (en) * | 2018-10-26 | 2021-01-05 | 陕西步长制药有限公司 | Detection method for determining content of effective components in leech capsule by HPLC-QQQ/MS method |
| CN110221009B (en) * | 2019-05-31 | 2020-04-17 | 北京澳合药物研究院有限公司 | Method for measuring butylphthalide related substances by HPLC |
| CN111413450B (en) * | 2020-05-20 | 2023-06-30 | 陕西国际商贸学院 | Detection method for radix astragali contained in traditional Chinese medicine preparation |
| CN111999395A (en) * | 2020-06-15 | 2020-11-27 | 陕西步长制药有限公司 | Fingerprint detection method of medicinal preparation |
| US20230213487A1 (en) * | 2020-06-15 | 2023-07-06 | Shaanxi Buchang Pharmaceutical Co., Ltd | Fingerprint Detection Method for Pharmaceutical Preparation |
| CN115184490B (en) * | 2022-06-28 | 2023-10-24 | 鲁南制药集团股份有限公司 | Establishment method and application of HPLC standard fingerprint of Hirudo vein relaxing capsule |
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| CN103207255B (en) * | 2013-03-12 | 2016-08-03 | 陕西步长制药有限公司 | A kind of detection method of content of NAOXINTONG JIAONANG |
| CN103411893B (en) * | 2013-07-29 | 2015-06-10 | 陕西步长制药有限公司 | Detection method for near infrared spectrum of Naoxintong capsule |
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