CN1621832A - Process and use of natural medicament high speed counter flow fingerprint spectrum - Google Patents
Process and use of natural medicament high speed counter flow fingerprint spectrum Download PDFInfo
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Abstract
The present invention relates to high speed countercurrent fingerprint method of natural medicine and its application. The present invention includes high speed countercurrent separating purification of natural medicine, establishing high speed countercurrent digital fingerprint method error, determining high speed countercurrent digital fingerprint method, and quantifying the difference degree of corresponding components in different samples with relative retention time and relative peak arean as main parameters as well as relative deviation, relative standard deviation and difference rate. The present invention may be used in the medicine quality identification in high precision, is suitable for analysis of high viscosity and adsorptive sample, and has high performance/cost ratio.
Description
Technical field
The present invention relates to the method for building up and the application of natural drug dactylogram, particularly relate to the method and the application of a kind of natural drug high-speed counter-current (HSCCC) dactylogram.
Technical background
From natural medicinal material, comprise Chinese medicine, mostly being artificial dispersion gathers, processes, be subjected to weather, regional difference and artifical influence factor very big, the standardization of raw material and semi-manufacture and end product quality, standardized management gap are bigger, for the homogeneous that guarantees product quality and stable, need to set up strict quality monitoring standard and good method for supervising.General analytical technology can only characterize the part chemical information, for example comprises not principal component of known active component, known non-active ingredient, a part.Know that now the drug effect of natural drug is not from any single active component, but by the various active composition, even relevant with synergy or " the giving birth to the gram effect " of " non-active ingredient ", so the chemical information of natural drug has certain ambiguity.
The quality assessment of natural drug commonly used is difficult point and the emphasis problem in research and the application always, though facts have proved of the existing more than one thousand years practical application of the value of natural drug, but in today of modern science and technology develop rapidly, how to estimate the quality of medicinal material effectively, how to guarantee the curative effect of natural drug, being one more and more needs the urgent problem that solves.Along with China enters WTO, as the world of medicine natural drug with independent intellectual property right or tcm product few in number, how to be admitted and to accept by the world, the quality of how to evaluate medicinal material could guarantee curative effect, just can accomplish to integrate with, become the important topic that medicinal industry faces with world level.Abroad the understanding to natural drug or Chinese medicine is based on the chemical constitution that it contained basically, also is the foundation of quality control.In recent years, the foreign study personnel adopt means such as TLC and HPLC, work up the TLC and the HPLC finger-print of the medicinal material of separate sources, for quality evaluation provides practicable scientific basis.The method of setting up the natural drug dactylogram is of a great variety, and the chromatographia method is main flow, especially TLC, HPLC and GC chromatographic technique.The natural drug finger-print not only possesses individual absolute uniqueness, also possess the uniqueness of species feature and the similarity between the individuality of the same race, the quality control index that makes medicine rises to detection to whole natural drug interior quality from original mensuration to certain several component content, the interior quality that can show natural drug to greatest extent, although for a certain concrete composition of medicinal material be what we may be still unclear, but can be with this important content as control and measurement quality of medicinal material standard, and make natural drug or traditional Chinese medicine research more meet the organic conception of traditional Chinese medicine, improved the good and bad standard of differentiating of the natural drug true and false.
Chang Yong chromatographic technique comprises the most: (1) thin layer chromatography (TLC): easy and simple to handle, no matter the crude drug compound has or not uv absorption or volatility, all can carry out qualitatively with TLC, but this method precision, accuracy and reappearance are relatively poor; (2) high performance liquid chromatography (HPLC): be applicable to crude drug compound of different nature, can analyze/separating bio alkali, compound and precision height such as glucoside, anthraquinone, organic acid, phenols, lactone, favorable reproducibility; But this method can't analysis of viscosity higher and easily cause the stationary phase adsorption sample, and instrument and consumptive material costliness are unfavorable for promoting; (3) gas chromatography (GC): it is very high to analyze usefulness, but restricted application, only is used for alkaloid, fat, lactone, phenol, sugar, animal kind medicine thing behind volatile ingredient and the derivatization.
High speed adverse current chromatogram (HSCCC) is a kind of liquid-liquid distribution chromatography that does not have solid phase carrier, has avoided post absorption and post to pollute; Simple to operate, system is changed easy; The accuracy height, favorable reproducibility; Have wide range of applications, be specially adapted to high viscosity and easily make stationary phase absorption material analysis with separate; The cost performance height suits to promote the use of in the crude drug place of production and economically underdeveloped area, is subjected to the problem of instrument condition restriction in helping to solve present standard formulation and implement; The HSCCC chromatogram can comprise abundant chemical information.
In addition, because adopting the chromatographic fingerprinting method to set up a difficult problem that the quality of medicinal material standard faces is need overcome general stability and the reappearance fluctuation that the various influence factors in the chromatographic run produce and be difficult to problem effective, easy, express-analysis figure spectrum signature, has proposed digitizing chromatographic fingerprint spectrum.Digitizing chromatographic fingerprint spectrum is a kind of new method at the complex component sample analysis, new technology, but it is not to be the improvement of chromatographic apparatus or chromatographic technique itself, but resulting chromatogram of stratographic analysis and chromatographic data are carried out suitable processing, complicated difficulty is distinguished that collection of illustrative plates transfers numeral intuitively to, set up series of parameters and corresponding a whole set of comparison rule and computing formula, make the analysis of complex component sample, evaluation become possibility.Therefore, digitizing chromatographic fingerprint spectrum is a kind of with the digitized technology of chromatogram, because digitized chromatographic fingerprint spectrum has been improved the floatability of original chromatogram effectively, make the comparison of relatively changing between the data directly perceived between the chromatogram, thereby strengthened the mutual comparability of sample.Digitizing chromatographic fingerprint spectrum not only can provide the qualitative description of complex component sample, also can set up its corresponding quantization index, makes more science, more objective of qualification result, also is easy to set up and use fingerprint spectrum library.
Existing digitizing dactylogram is just listed in the relative retention time and the relative peak area of different batches (place of production) each corresponding component of sample in two tables.For example the difference degree (table 1) of the relative retention time of sample 1 and sample N does not adopt relative deviation, relative standard deviation etc. to carry out clear and definite calculating; The difference (table 2) of the relative peak area between any 2 samples also not by variance rate be embodied in the table of digital finger-print spectrum (Hong Xiaokun, Wang Zhihua. Chinese medicine digitizing chromatographic fingerprint spectrum. Shanghai: the .2003. of Shanghai science tech publishing house).
The relative retention time of table 1 different batches (place of production) each corresponding component of sample
| Batch | Elution fraction | |||||||
| ????1 | ???1 | ???2 | ???3 | ???I | ???N | |||
| ????2 | ||||||||
| ????3 | ||||||||
| ????I | ||||||||
| ????N | ||||||||
The relative peak area of table 2 different batches (place of production) each corresponding component of sample
| Batch | Elution fraction | |||||||
| ????1 | ???1 | ???2 | ???3 | ???I | ???N | |||
| ????2 | ||||||||
| ????3 | ||||||||
| ????I | ||||||||
| ????N | ||||||||
Summary of the invention
The technical problem to be solved in the present invention is to overcome existing method when the natural drug dactylogram is set up, the quantification of different batches sample is the defective of intuitive difference relatively, and a kind of method that is applicable to high-speed counter-current (HSCCC) dactylogram of high viscosity and easy adsorbent is provided.
Technical scheme of the present invention comprises the steps:
(1) choose with reference to the peak:
With reference to the peak method, must meet following condition: separate well with adjacent chromatographic peak, peak position is placed in the middle in adopting; Be respectively to wait to reflect the total chromatographic peak of sample; It is the important indicator composition of this kind.If analysis time is long and the peak number order is more, can select twoly with reference to the peak, promptly in the first half chromatographic peak, select mark peak one in, in the latter half chromatographic peak, select again and mark the peak in one, form two relative retention value series foundation as a comparison.Relative retention time is retention time and interior recently the quantizing of using chromatographic peak with reference to the peak retention time.Relative peak area adopts with reference to the p-ratio method: as benchmark, making comparisons in the peak area of other each chromatographic peaks and it, obtains a series of area ratio thus with interior peak area with reference to the peak.
(2) foundation of method error
At first determine the reappearance of method, same batch sample is carried out replica test, determine the error of method, the frequency n of revision test should be determined according to mathematical statistics method.Method error comprises as the relative deviation of the relative retention time of qualitative foundation and relative standard deviation, and as the relative deviation and the relative standard deviation of the relative peak area of quantitative basis.If in the method error scope, this group peak of decidable is same peak (component); Then can be considered different component (table 3) if exceed method error.
The method error of table 3 HSCCC digitizing dactylogram
| ???1 | ???2 | ???3 | ????i | ???n | ||||||||||
| Relative t R | Experiment 1 | |||||||||||||
| Experiment i | ||||||||||||||
| Experiment n | ||||||||||||||
| On average | ||||||||||||||
| Relative t
R | Experiment | 1 | ||||||||||||
| Experiment i | ||||||||||||||
| Experiment n | ||||||||||||||
| Relative t R????RSD(％) | ||||||||||||||
| Relative A | | |||||||||||||
| Experiment i | ||||||||||||||
| Experiment n | ||||||||||||||
| On average | ||||||||||||||
| Relative A | Experiment | 1 | ||||||||||||
| Experiment I | ||||||||||||||
| Experiment n | ||||||||||||||
| Relative A RSD (%) | ||||||||||||||
(3) make up HSCCC digital finger-print spectrum
Digitizing dactylogram (table 4) is to be based upon on the experimental result basis of chromatographic fingerprinting.When making up the digitizing dactylogram, at first selected with reference to peak i, generally follow following principle: the one, choose the stronger peak in whole chromatogram centre position, the 2nd, better with the degree of separation at two peaks, front and back, the 3rd, this peak is known compound as far as possible, and is important effective constituent.
The calculating of relative retention time is that the retention time with peak i is 1 in the table 4, and with the retention time at other peak by comparison, so the retention time at other peaks all is greater than or less than 1.Mean value is meant the relative retention time t at correspondence peak in the table 4
RMean value.Relative retention time t
RRelative standard deviation and relative standard deviation calculate according to following formula.
Relative peak area A adopts the normalized area value in the table 4, promptly the area with reference peak i is 1, with other chromatographic peak areas by comparison, obtain a series of area ratio, variance rate for the same peak of different batches sample is calculated, if determining a certain batch is standard, can compare calculating with it, all not settling the standard as if all batches can be more definite with mean value.
Characteristic fingerprint peak variance rate=| A
I Mark-A
I Mirror|/A
I Mark
The component of relative retention time in the method error scope is placed on a window, promptly thinks to be same component in the method error scope, what exceed thinks different component.
Variance rate is meant each fingerprint peaks and its respective standard fingerprint peaks difference degree between the two.The total variances rate then is the variance rate summation with each fingerprint peaks in the sample.The total variances rate is big more, and expression relatively both sides differs big more, has concluded the comprehensive parameter of qualitative evaluation and quantificational description two aspect information simultaneously.
Characteristic fingerprint peak variance rate=| A
I Mark-A
I Mirror|/A
I Mark
The relative retention time, the relative peak area that have not only comprised different samples in HSCCC digitizing dactylogram simultaneously are also with the difference degree of corresponding component in relative deviation, relative standard deviation and the variance rate different samples that come quantificational description.
(table 4) HSCCC digitizing dactylogram
| Elution fraction | |||||||||||||
| ???1 | ???2 | ???3 | ????i | ???n | |||||||||
| Relative | Batch | 1 | |||||||||||
| Batch i | |||||||||||||
| Batch n | |||||||||||||
| On average | |||||||||||||
| Relative t
R | Batch | 1 | |||||||||||
| Batch i | |||||||||||||
| Batch n | |||||||||||||
| Relative t R?RSD (％) | |||||||||||||
| Relative peak area | Batch 1 | ||||||||||||
| Batch i | |||||||||||||
| Batch n | |||||||||||||
| | Batch | 1 | |||||||||||
| Batch n | |||||||||||||
(4) characteristic fingerprint
The general character of a class natural drug sample has been represented at total peak, also is the characteristic of this kind, is and other kind difference parts, thereby has constituted the peculiar fingerprint of this biosome, is called characteristic fingerprint.The peak group that characteristic fingerprint is made up of a series of characteristic fingerprint peak, characteristic fingerprint can be used to contain the discriminating between the different natural drugs of identical effective constituent, is mainly used in kind and differentiates.The value difference of fingerprint peaks has reflected the difference of same breed organism, the different place of production, Different Harvesting Time quality, thereby can be used as the Quality Control foundation of raw material and patent medicine again.
The characteristic fingerprint peak must be the common peak of same kind sample, and has been identified as the peak that this medicinal material is imitated composition or special composition.The characteristic fingerprint peak should account for more than 60% of chromatographic peak sum.
The characteristic fingerprint curve is made of characteristic fingerprint and the corresponding area normalization value of each fingerprint peaks, and the situation of certain natural drug characteristic fingerprint is described in the mode of curve map.It is a kind of audio-visual picture, simultaneously the quality and quantity at each characteristic fingerprint peak is shown with the form of curve.Can will wait to reflect in the same coordinate system system characteristic fingerprint peak of medicine materical crude slice and respective standard shows simultaneously, can be clear that both similarity degrees.
The invention provides a kind of method of natural drug dactylogram, different content natural drug from various technology approach preparations, adopt high-speed counter-current chromatograph (HSCCC) can directly advance slightly get sample in a large number product or synthetic mixture, separating resulting can reach quite high purity.High-speed countercurrent chromatography can avoid post absorption and post to pollute, be specially adapted to high viscosity and easily cause the analysis of the sample of stationary phase absorption, its cost performance height helps applying in the crude drug place of production and economically underdeveloped area, is the important supplement to existing fingerprint spectrum method.
The invention provides a kind of application of natural drug dactylogram, the digitizing dactylogram is not only used the notion of relative value, and the finger-print that has solved graphic form is subjected to operation factors to influence, be difficult to carry out multiple batches of quantification problem relatively.And it is clear and definite on the basis of method error, set up series of parameters and corresponding a whole set of comparison rule and computing formula, the relative deviation and the relative standard deviation of relative retention time in the digital finger-print spectrum, have directly been introduced, and the variance rate of relative peak area, with intuitively, show the difference degree of each corresponding component in the different samples quantitatively, realized between any 2 samples quantification intuitively relatively.Can improve the good and bad standard of differentiating of the medicinal material true and false, as the important content of control and measurement natural drug quality standard.
Description of drawings
Fig. 1-the 1st, the HSCCC collection of illustrative plates of the red sage root is produced in Hebei
Fig. 1-2 is the HSCCC collection of illustrative plates that the red sage root is produced in Shandong
Fig. 1-the 3rd, the HSCCC collection of illustrative plates of the red sage root is produced in Jiangsu
Fig. 2 is that the feature of red sage root quinones refers to curve
Fig. 3 is that the feature of root of red-rooted salvia phenolic acid compounds refers to curve
Fig. 4 is that the feature of saussurea involucrata flavone compound refers to curve
Fig. 5 is that the feature of earthworm alkaloid compound refers to curve
Embodiment
Formulate the HSCCC digitizing dactylogram of salvia piece
1. instrument and reagent
(1) instrument:
1) TBE-300 half countercurrent chromatography instrument (Shenzhen is with field biochemical technology company limited, China).Polyfluortetraethylene pipe is wrapped in and forms spiral pipe (pipe diameter 2.6mm, separated volume 300mL), sample introduction circle 20mL on 3 transverse axis.High speed adverse current chromatogram has been equipped with ram type pump (S series, the logical technological development Ltd of the holy benefit in Beijing, China), Ultraviolet Detector (8823A, Beijing new technology application research institute, China), registering instrument (3057 types, Sichuan register instrument factory, China).
2) high performance liquid chromatography (10Avp series, Tianjin, island, Japan): double pump system (LC-10ATvp), Ultraviolet Detector (SPD-10Avp), column oven (CTO-10ASvp), system controller (SCL-10Avp), Control Software (Class-vp), 20 μ l sample introduction circles; Chromatographic column: reverse C
18Post (150mm * 4.6mm I.D., 5 μ m).
(2) reagent:
1) high speed adverse current chromatogram reagent adopts and analyzes pure (Tianjin chemical reagent one factory).Water is by counter-infiltration Mili-Q (18M Ω, Milipore, USA) preparation.
2) reagents for high performance liquid chromatography adopt the one-level chromatographically pure (Fisher Scientific Co.Ltd., UK).(18M Ω, Milipore is USA) by using behind the 0.45 μ m filter membrane filtration under diminished pressure for the water that reverse-osmosis treated is crossed.Standard reference material is mixed with 1mg/mL with the moving phase of high speed adverse current chromatogram, uses through 0.45 μ m membrane filtration.
(3) salvia piece: from 3 places of production: Hebei, Shandong, Jiangsu.
(4) standard reference material: National Drug Administration, Chinese medicine and biological products assay institute provide.
2. the digitizing dactylogram is set up
Figure 1 shows that the HSCCC collection of illustrative plates of the red sage root in 3 places of production.As seen from the figure, through the HSCCC purifying, the red sage root in 3 places of production respectively obtains 12 eluting peaks, the number unanimity at peak.Each eluting peak is analyzed: measure the retention time of each eluting peak on reversed-phase high-performance liquid chromatography (HPLC); In ultraviolet-visible spectrophotometer 900-200nm scope each eluting peak is carried out full wavelength scanner, spectrum is absorbed.Can judge that from HPLC retention time and ultra-violet absorption spectrum the individual eluting peak of n (n is 3-12) in 3 place of production red sage root elution curves is same component.That is to say, the HSCCC wash-out collection of illustrative plates of 3 place of production reds sage root, not only eluting peak number unanimity, and corresponding eluting peak is same component.
(1) chooses eluting peak 7 for calculating relative retention time and relative peak area, avoid the data fluctuations that causes because of factors such as instrumentations with reference to the peak.
(2) method error of calculating HSCCC digitizing dactylogram
The method error of the HSCCC digitizing dactylogram of salvia piece
| ???1 | ???2 | ????3 | ???4 | ???5 | ???6 | ???7 | ???8 | ???9 | ???10 | ????11 | ????12 | ||
| Relative t R | Experiment 1 | ???0.30 | ???0.34 | ????0.39 | ???0.49 | ???0.64 | ???0.79 | ???1 | ???1.55 | ???2.02 | ???2.62 | ????3.53 | ????4.14 |
| Experiment 2 | ???0.29 | ???0.33 | ????0.40 | ???0.49 | ???0.64 | ???0.78 | ???1 | ???1.52 | ???1.97 | ???2.57 | ????3.43 | ????4.02 | |
| Experiment 3 | ???0.29 | ???0.34 | ????0.39 | ???0.49 | ???0.63 | ???0.77 | ???1 | ???1.57 | ???2.07 | ???2.64 | ????3.63 | ????4.24 | |
| On average | ???0.293 | ???0.337 | ????0.393 | ???0.490 | ???0.637 | ???0.780 | ???1 | ???1.547 | ???2.020 | ???2.610 | ????3.530 | ????4.133 | |
| Relative t RRelative deviation | Experiment 1 | ???0.02 | ???0.01 | ????-0.01 | ???0 | ???0.01 | ???0.01 | ???0 | ???0.01 | ???0 | ???0.01 | ????0 | ????0.01 |
| Experiment 2 | ???-0.01 | ???-0.02 | ????0.02 | ???0 | ???0.01 | ???0 | ???0 | ???-0.02 | ???-0.02 | ???-0.02 | ????-0.03 | ????-0.03 | |
| Experiment 3 | ???-0.01 | ???0.01 | ????-0.01 | ???0 | ???-0.01 | ???-0.01 | ???0 | ???0.01 | ???0.02 | ???0.01 | ????0.03 | ????0.03 | |
| Relative t R??RSD(％) | ???0.02 | ???0.02 | ????0.02 | ???0 | ???0.01 | ???0.01 | ???0 | ???0.02 | ???2.47 | ???1.38 | ????2.83 | ????2.16 | |
| Relative A | Experiment 1 | ???0.17 | ???0.10 | ????0.16 | ???0.33 | ???0.29 | ???0.31 | ???1 | ???1.50 | ???0.76 | ???1.04 | ????4.56 | ????0.37 |
| Experiment 2 | ???0.17 | ???0.10 | ????0.16 | ???0.32 | ???0.30 | ???0.31 | ???1 | ???1.51 | ???0.71 | ???1.01 | ????4.25 | ????0.35 | |
| Experiment 3 | ???0.15 | ???0.10 | ????0.16 | ???0.29 | ???0.29 | ???0.29 | ???1 | ???1.45 | ???0.73 | ???0.95 | ????4.33 | ????0.33 | |
| On average | ???0.163 | ???0.100 | ????0.160 | ???0.313 | ???0.293 | ???0.303 | ???1 | ???1.487 | ???0.733 | ???1.000 | ????4.380 | ????0.35 | |
| Relative A relative deviation | Experiment 1 | ???0.04 | ???0 | ????0 | ???0.05 | ???-0.01 | ???0.02 | ???0 | ???-0.01 | ???0.04 | ???0.04 | ????0.04 | ????0.06 |
| Experiment 2 | ???0.04 | ???0 | ????0 | ???0.02 | ???0.02 | ???0.02 | ???0 | ???0.02 | ???-0.03 | ???0.01 | ????-0.03 | ????0 | |
| Experiment 3 | ???-0.92 | ???0 | ????0 | ???-0.07 | ???-0.01 | ???-0.04 | ???0 | ???-0.02 | ???-0.01 | ???-0.05 | ????-0.01 | ????-0.06 | |
| Relative A RSD (%) | ???7.09 | ???0 | ????0 | ???6.65 | ???1.69 | ???1.63 | ???0 | ???2.16 | ???3.43 | ???4.58 | ????3.67 | ????5.71 | |
Method error comprises two aspects: the error range of (1) relative retention time: the relative standard deviation of relative retention time is less than 3%; (2) error range of relative peak area: the relative standard deviation of relative peak area is less than 8%.That is: the RSD of relative retention time＜3% is thought same component; The RSD of relative peak area＜8% thinks that content is identical.
On this basis, relative retention time is placed under the window difference degree of each corresponding component in the more different samples in the method error scope.
(3) make up salvia piece HSCCC digitizing dactylogram
The HSCCC digitizing dactylogram of salvia piece
| ??1 | ???2 | ???3 | ??4 | ??5 | ??6 | ??7 | ??8 | ??9 | ??10 | ??11 | ??12 | ||
| Relative t R | Produce in Hebei | ??0.41 | ???0.47 | ???0.54 | ??0.61 | ??0.72 | ??0.84 | ??1.00 | ??1.38 | ??1.64 | ??2.11 | ??3.04 | ??3.62 |
| Produce in Shandong | ??0.40 | ???0.47 | ???0.53 | ??0.61 | ??0.73 | ??0.84 | ??1.00 | ??1.37 | ??1.64 | ??2.10 | ??3.07 | ??3.63 | |
| Produce in Jiangsu | ??0.40 | ???0.49 | ???0.56 | ??0.63 | ??0.73 | ??0.86 | ??1.00 | ??1.35 | ??1.64 | ??2.07 | ??3.10 | ??3.65 | |
| On average | ??0.40 ??3 | ???0.47 ???7 | ???0.54 ???3 | ??0.61 ??3 | ??0.72 ??7 | ??0.84 ??7 | ??1.00 ??0 | ??1.36 ??7 | ??1.64 | ??2.09 ??3 | ??3.07 ??0 | ??3.63 ??3 | |
| Relative t RRelative deviation | Produce in Hebei | ??1.7 | ???-1.5 | ???-0.6 | ??-0.5 | ??-1.0 | ??-0.8 | ??0 | ??1.0 | ??0 | ??0.8 | ??-1.0 | ??-0.4 |
| Produce in Shandong | ??-0.7 | ???-1.5 | ???-2.4 | ??-0.5 | ??0.4 | ??-0.8 | ??0 | ??0.2 | ??0 | ??0.3 | ??0 | ??-0.0 ??8 | |
| Produce in Jiangsu | ??-0.7 | ???2.7 | ???3.1 | ??2.8 | ??0.4 | ??1.5 | ??0 | ??-1.2 | ??0 | ??-1.1 | ??1.0 | ??0.5 | |
| Relative t R????RSD(％) | ??1.44 | ???2.42 | ???2.81 | ??1.87 | ??0.8 | ??1.36 | ??0 | ??1.11 | ??0 | ??1 | ??1 | ??0.42 | |
| Relative peak area | Produce in Hebei | ??0.10 | ???0.01 | ???0.02 | ??0.10 | ??0.12 | ??0.03 | ??1.00 | ??0.27 | ??0.46 | ??0.54 | ??2.05 | ??0.42 |
| Produce in Shandong | ??0.22 | ???0.01 | ???0.01 | ??0.19 | ??0.08 | ??0.07 | ??1.00 | ??1.56 | ??0.13 | ??0.62 | ??3.77 | ??0.10 | |
| Produce in Jiangsu | ??0.09 | ???0.01 | ???0.01 | ??0.15 | ??0.13 | ??0.02 | ??1.00 | ??0.59 | ??0.28 | ??0.89 | ??3.17 | ??0.21 | |
| Relative A RSD (%) | ??52.8 ??1 | ???0 | ???44.5 ???2 | ??30.6 ??8 | ??24.0 ??5 | ??24.0 ??5 | ??0 | ??83.2 ??4 | ??56.9 ??7 | ??53.8 ??3 | ??29.3 ??1 | ??66.9 ??1 | |
| Variance rate | Produce in Shandong | ??0.40 | ???0.67 | ???0.8 | ??0.25 | ??0.58 | ??0.8 | ??0.34 | ??2.82 | ??0.82 | ??0.25 | ??0.22 | ??0.84 |
| Produce in Jiangsu | ??0.30 | ???0.67 | ???0.80 | ??0.15 | ??0.21 | ??0.40 | ??0.22 | ??0.72 | ??0.53 | ??0.28 | ??0.21 | ??0.60 | |
(4) draw the characteristic fingerprint curve
There are not non-total peak, peak 1 in the HSCCC dactylogram of red sage root fat-soluble compound
#～12
#All be that these three samples are total, can constitute the characteristic fingerprint of HSCCC, draw characteristic fingerprint curve (Fig. 2) with these 12 peaks as the characteristic fingerprint peak.
Formulate the HSCCC digitizing dactylogram of root of red-rooted salvia phenolic acid compounds
1. instrument device and reagent: TBE-300 half countercurrent chromatography instrument (Tongtian Biochemical Technology Co., Ltd., Shanghai, China).Polyfluortetraethylene pipe is wrapped in and forms spiral pipe (pipe diameter 2.6mm, separated volume 300mL), sample introduction circle 20mL on 3 transverse axis.(be the distance between the supporting axis and the axis of centres, R) be 5cm, the β value is from 0.5～0.8 (β=r/R, r are meant the distance of spiral pipe to supporting axis to separation semidiameter).High speed adverse current chromatogram has been equipped with constant flow pump (S series, Beijing logical technological development Ltd of holy benefit, China), Ultraviolet Detector (8823A, Beijing new technology application research institute, China), registering instrument (3057 types, Sichuan register instrument factory, China) and data acquisition system (DAS) (N2000, intellectual technology research institute of Zhejiang University).Normal hexane, ethyl acetate, methyl alcohol all adopt analysis pure.The rough sample of different batches salvianolic acid is given birth to company available from the Xi'an letter, directly goes up sample with moving phase dissolving back.
2. method: the system of normal hexane-methanol-water=1.5: 1.5: 5 below is a moving phase mutually, pH2.5～2.6,850r/min, 2mL/min, whole system is set up mobile equilibrium, inject a certain batch of medicinal material crude extract from sampling valve, each component is separated by its partition factor in two-phase; Use UV-detector to detect eluent, draw the uv absorption of each component and the graph of a relation of elution time; 3 batches of samples all obtain 11 of wash-outs through the elution curve of high speed adverse current chromatogram separation and purification.
3. dactylogram preparation method: according to method error: RSD＜3% to retention time is thought same component; The RSD of relative peak area＜8% thinks that content is identical, and relative retention time is placed under the window difference degree of each corresponding component in the more different samples in the method error scope.The retention time and the peak area of each component of different batches sample are listed in the table, and calculated the main body that relative deviation, relative standard deviation and variance rate constitute HSCCC digitizing dactylogram.
The HSCCC digitizing dactylogram of root of red-rooted salvia phenolic acid compounds
| ??1 | ??2 | ??3 | ??4 | ??5 | ??6 | ??7 | ??8 | ??9 | ??10 | ??11 | ||
| Relative retention time | Produce in Hebei | ??0.90 | ??0.91 | ??0.92 | ??0.94 | ??0.96 | ??1 | ??1.01 | ??1.05 | ??1.06 | ??1.13 | ??1.16 |
| Produce in Shandong | ??0.90 | ??0.91 | ??0.92 | ??0.94 | ??0.96 | ??1 | ??1.02 | ??1.04 | ??1.06 | ??1.12 | ??1.15 | |
| Produce in Jiangsu | ??0.90 | ??0.91 | ??0.92 | ??0.94 | ??0.96 | ??1 | ??1.01 | ??1.04 | ??1.06 | ??1.12 | ??1.15 | |
| On average | ??0.900 | ??0.910 | ??0.920 | ??0.940 | ??0.960 | ??1 | ??1.013 | ??1.043 | ??1.060 | ??1.123 | ??1.153 | |
| Relative t RRelative deviation | Produce in Hebei | ??0 | ??0 | ??0 | ??0 | ??0 | ??0 | ??-0.3 | ??0.7 | ??0 | ??0.6 | ??0.6 |
| Produce in Shandong | ??0 | ??0 | ??0 | ??0 | ??0 | ??0 | ??0.7 | ??-0.3 | ??0 | ??-0.3 | ??-0.3 | |
| Produce in Jiangsu | ??0 | ??0 | ??0 | ??0 | ??0 | ??0 | ??-0.3 | ??-0.3 | ??0 | ??-0.3 | ??-0.3 | |
| Relative t R?RSD(％) | ??0 | ??0 | ??0 | ??0 | ??0 | ??0 | ??0.57 | ??0.55 | ??0 | ??0.52 | ??0.50 | |
| Relative peak area | Produce in Hebei | ??0.06 | ??0.03 | ??0.13 | ??0.17 | ??0.02 | ??1 | ??0.03 | ??0.31 | ??0.10 | ??1.11 | ??0.08 |
| Produce in Shandong | ??0.10 | ??0.03 | ??0.15 | ??0.10 | ??0.03 | ??1 | ??0.06 | ??0.12 | ??0.08 | ??0.41 | ??0.08 | |
| Produce in Jiangsu | ??0.04 | ??0.08 | ??0.13 | ??0.20 | ??0.04 | ??1 | ??0.02 | ??1.01 | ??0.04 | ??2.05 | ??0.10 | |
| Relative A RSD (%) | ??45.61 | ??61.4 | ??8.4 | ??32.7 | ??33.3 | ??0 | ??57.3 | ??97.7 | ??41.9 | ??69.2 | ??13.3 | |
| Variance rate | Produce in Shandong | ??0.98 | ??0.27 | ??0.41 | ??0.26 | ??0.44 | ??0.24 | ??1.48 | ??0.52 | ??0.06 | ??0.54 | ??0.36 |
| Produce in Jiangsu | ??0.61 | ??0.86 | ??0.28 | ??0.17 | ??0.18 | ??0.29 | ??0.60 | ??1.29 | ??0.67 | ??0.31 | ??0.10 | |
Set up the characteristic fingerprint curve of HSCCC on this basis, because of not having non-total peak, peak 1 in its dactylogram
#～11
#All be that these three samples are total, can constitute the characteristic fingerprint of salvianolic acid HSCCC, draw characteristic fingerprint curve (Fig. 3) with these 11 peaks as the characteristic fingerprint peak.
Work out saussurea involucrata flavonoids high speed adverse current chromatogram digitizing dactylogram
1. instrument and reagent: TBE-300 half countercurrent chromatography instrument (Tongtian Biochemical Technology Co., Ltd., Shanghai).Polyfluortetraethylene pipe is wrapped in and forms spiral pipe (pipe diameter 2.6mm, separated volume 300mL), sample introduction circle 20mL on 3 transverse axis.(be the distance between the supporting axis and the axis of centres, R) be 5cm, the β value is from 0.5～0.8 (β=r/R, r are meant the distance of spiral pipe to supporting axis to separation semidiameter).High speed adverse current chromatogram has been equipped with constant flow pump (S series, Beijing logical technological development Ltd of holy benefit, China), Ultraviolet Detector (8823A, Beijing new technology application research institute, China), registering instrument (3057 types, Sichuan register instrument factory, China) and data acquisition system (DAS) (N2000, intellectual technology research institute of Zhejiang University).It is pure that chloroform, methyl alcohol are analysis.Herba Saussureae Involueratae is available from Xinjiang autonomous region.
2. method: the rough Preparation Method of saussurea involucrata flavone compound is as follows: dry lanatechead saussurea herb with flower was removed impurity weighing 20g, with the 70% ethanol room temperature lixiviate of 300mL 7 days; The concentrating under reduced pressure leaching liquor; Add equal-volume water, jolting is even, leaves standstill 1h; Centrifugal removal precipitation 1000g * 6min; Get the supernatant concentrating under reduced pressure and become medicinal extract; Get during HSCCC separates and go up sample after 80 μ L are dissolved in 3mL moving phase.The HSCCC separation condition: selecting chloroform-methanol-water for use is the solvent system of 9-12-8, below is moving phase mutually; Engine speed 800r/min, flow rate of mobile phase 1.7mL/min, whole system is set up mobile equilibrium, injects a certain batch of medicinal material crude extract from sampling valve, and each component is separated by its partition factor in two-phase; Use UV-detector to detect eluent, draw the uv absorption of each component and the graph of a relation of elution time; 3 batches of saussurea involucrata chromocor compound samples all obtain 7 of eluting peaks through the elution curve of high speed adverse current chromatogram separation and purification.
3. dactylogram preparation method: according to method error: RSD＜3% to retention time is thought same component; The RSD of relative peak area＜8% thinks that content is identical, and relative retention time is placed under the window difference degree of each corresponding component in the more different samples in the method error scope.
The retention time and the peak area of each component of different batches sample are listed in the table, and calculated the main body that relative deviation, relative standard deviation and variance rate constitute HSCCC digitizing dactylogram.
The HSCCC digitizing dactylogram of saussurea involucrata flavone compound
| ????1 | ????2 | ????3 | ????4 | ????5 | ????6 | ????7 | ||
| Relative retention time | Produce in Hebei | ????1.00 | ????1.04 | ????1.12 | ????1.13 | ????1.15 | ????1.75 | ????1.83 |
| Produce in Shandong | ????1.00 | ????1.04 | ????1.12 | ????1.14 | ????1.16 | ????1.81 | ????1.89 | |
| Produce in Jiangsu | ????1.00 | ????1.04 | ????1.11 | ????1.13 | ????1.15 | ????1.74 | ????1.80 | |
| On average | ????1.000 | ????1.040 | ????1.117 | ????1.133 | ????1.153 | ????1.767 | ????1.840 | |
| Relative t RRelative deviation | Produce in Hebei | ????0 | ????0 | ????0.3 | ????-0.3 | ????-0.3 | ????-1.0 | ????-0.5 |
| Produce in Shandong | ????0 | ????0 | ????0.3 | ????0.6 | ????0.6 | ????2.4 | ????2.7 | |
| Produce in Jiangsu | ????0 | ????0 | ????-0.6 | ????-0.3 | ????-0.3 | ????-1.5 | ????-2.2 | |
| Relative t R?RSD(％) | ????0 | ????0 | ????0.52 | ????0.51 | ????0.50 | ????2.14 | ????2.49 | |
| Relative peak area | Produce in Hebei | ????1.00 | ????0.06 | ????2.08 | ????0.39 | ????0.14 | ????0.21 | ????0.09 |
| Produce in Shandong | ????1.00 | ????0.11 | ????1.98 | ????0.72 | ????0.52 | ????0.13 | ????0.07 | |
| Produce in Jiangsu | ????1.00 | ????0.08 | ????2.57 | ????0.77 | ????0.51 | ????0.21 | ????0.08 | |
| Relative A RSD (%) | ????0 | ????30.3 | ????14.3 | ????32.8 | ????55.5 | ????25.8 | ????12.5 | |
| Variance rate | Produce in Shandong | ????1.43 | ????3.25 | ????1.31 | ????3.53 | ????8.41 | ????0.53 | ????1 |
| Produce in Jiangsu | ????0.72 | ????1.00 | ????1.30 | ????2.43 | ????5.54 | ????0.73 | ????0.52 | |
Set up the characteristic fingerprint curve of HSCCC on this basis, because of not having non-total peak, peak 1 in its dactylogram
#～7
#All be that these three samples are total, can constitute the characteristic fingerprint of HSCCC, draw characteristic fingerprint curve (Fig. 4) with these 7 peaks as the characteristic fingerprint peak.
The HSCCC digitizing dactylogram of earthworm alkaloid compound
1. instrument and reagent:
TBE-300 half countercurrent chromatography instrument (Tongtian Biochemical Technology Co., Ltd., Shanghai, China).Polyfluortetraethylene pipe is wrapped in and forms spiral pipe (pipe diameter 2.6mm, separated volume 300mL), sample introduction circle 20mL on 3 transverse axis.(be the distance between the supporting axis and the axis of centres, R) be 5cm, the β value is from 0.5～0.8 (β=r/R, r are meant the distance of spiral pipe to supporting axis to separation semidiameter).High speed adverse current chromatogram has been equipped with constant flow pump (S series, Beijing logical technological development Ltd of holy benefit, China), Ultraviolet Detector (8823A, Beijing new technology application research institute, China), registering instrument (3057 types, Sichuan register instrument factory, China) and data acquisition system (DAS) (N2000, intellectual technology research institute of Zhejiang University).Each organic solvent all adopts analysis pure.
2. method:
The preparation method of earthworm leachate is as follows: gather the earthworm 10g that lives; Clean the body surface dirt with distilled water; Put and deposit 3h in the distilled water, allow it drain voluntarily,, make the gastrovascular cavity emptying through handling for several times; Each 5g places in the glassware with clean worm, adds sucrose 5g liquid in the body is deviate from.The HSCCC separation condition: chloroform-methanol-water=4-3-2 system below is moving phase mutually; HSCCC engine speed 850r/min, the whole system of flow rate of mobile phase 2mL/min is set up mobile equilibrium, injects a certain batch of medicinal material crude extract from sampling valve, and each component is separated by its partition factor in two-phase; Detect eluent with UV-detector, draw the uv absorption of each component and the graph of a relation of elution time; 3 batches of earthworm alkaloid compound samples all obtain 8 of eluting peaks through the elution curve of separation and purification.
3. dactylogram preparation method: according to method error: RSD＜3% to retention time is thought same component; The RSD of relative peak area＜8% thinks that content is identical, and relative retention time is placed under the window difference degree of each corresponding component in the more different samples in the method error scope.
The retention time and the peak area of each component of different batches sample are listed in the table, and calculated the main body that relative deviation, relative standard deviation and variance rate constitute HSCCC digitizing dactylogram.
The HSCCC digitizing dactylogram of earthworm alkaloid compound
| ????1 | ????2 | ????3 | ????4 | ????5 | ????6 | ????7 | ????8 | ||
| Relative retention time | Produce in Hebei | ????0.55 | ????0.94 | ????1 | ????1.16 | ????1.39 | ????1.56 | ????1.63 | ????1.93 |
| Produce in Shandong | ????0.50 | ????0.93 | ????1 | ????1.15 | ????1.37 | ????1.52 | ????1.60 | ????1.89 | |
| Produce in Jiangsu | ????0.48 | ????0.94 | ????1 | ????1.14 | ????1.36 | ????1.50 | ????1.56 | ????1.82 | |
| On average | ????0.510 | ????0.937 | ????1 | ????1.150 | ????1.373 | ????1.527 | ????1.600 | ????1.880 | |
| Relative t RRelative deviation | Produce in Hebei | ????7.8 | ????0.3 | ????0 | ????0.9 | ????1.2 | ????2.2 | ????1.9 | ????2.7 |
| Produce in Shandong | ????-2.0 | ????-0.7 | ????0 | ????0 | ????-0.2 | ????-0.5 | ????0 | ????0.5 | |
| Produce in Jiangsu | ????-5.9 | ????0.3 | ????0 | ????-0.9 | ????-0.9 | ????-1.8 | ????-2.5 | ????-3.2 | |
| Relative t R?RSD(％) | ????7.07 | ????0.62 | ????0 | ????0.87 | ????1.11 | ????1.67 | ????2.21 | ????2.97 | |
| Relative peak area | Produce in Hebei | ????0.71 | ????0.29 | ????1 | ????0.30 | ????0.74 | ????0.35 | ????1.03 | ????0.32 |
| Produce in Shandong | ????1.86 | ????0.32 | ????1 | ????0.48 | ????1.31 | ????0.41 | ????1.57 | ????0.72 | |
| Produce in Jiangsu | ????0.22 | ????0.24 | ????1 | ????0.29 | ????0.65 | ????0.33 | ????1.24 | ????0.35 | |
| Relative A RSD (%) | ????90.3 | ????14.5 | ????0 | ????29.7 | ????39.8 | ????9.5 | ????21.2 | ????48.4 | |
| Variance rate | Produce in Shandong | ????0.40 | ????0.42 | ????0.47 | ????0.15 | ????0.06 | ????0.38 | ????0.19 | ????0.20 |
| Produce in Jiangsu | ????0.72 | ????0.27 | ????0.12 | ????0.14 | ????0.23 | ????0.18 | ????0.06 | ????0.03 | |
Set up the characteristic fingerprint curve of HSCCC on this basis, because of not having non-total peak, peak 1 in its dactylogram
#～8
#All be that these three samples are total, can constitute the characteristic fingerprint of HSCCC, draw characteristic fingerprint curve (Fig. 5) with these 8 peaks as the characteristic fingerprint peak.
Claims (9)
1, a kind of method of natural drug high speed counter flow fingerprint spectrum is characterized in that comprising the following steps:
(1) preparation of test sample: adopt routine techniques, the crude extract of preparation batch crude drug more than 3;
(2) determine the main compound type, choose dicyandiamide solution, regulate the ratio of each solvent in the dicyandiamide solution, choose the suitable system of partition factor; Arbitrary in the above-mentioned system is stationary phase mutually, and another is moving phase mutually, and each component is by its partition factor first separation purifying in two-phase;
(3) the online detection eluent of UV-detector is drawn the uv absorption of each component and the graph of a relation of elution time; Obtain the chromatogram of each batch medicinal material, choose total fingerprint peaks; Calculate the method error (table 3) of high speed adverse current chromatogram digitizing dactylogram; Formulate the digitizing dactylogram (table 4) of each batch medicinal material, be as the criterion, the calculated difference rate with the standard medicinal material;
The method error of (table 3) HSCCC digitizing dactylogram
(table 4) HSCCC digitizing dactylogram
2. the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 1, it is characterized in that: at 10～40 ℃, open high-speed counter-current chromatograph, adopt the main frame forward or reverse, engine speed is 750～950rpm, and with the flow velocity of 1.0～2.5mL/min moving phase is pumped in the separating column, whole system is set up mobile equilibrium.
3. the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 2, it is characterized in that: the dicyandiamide solution first that three components of salvia piece constitute is normal hexane-ethanol-water, volume ratio 10: 5.5: 4.5, dicyandiamide solution second normal hexane-ethanol-water volume ratio 10: 7: 3.
4. the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 2 is characterized in that: the salvianolic acid dicyandiamide solution is normal hexane-methyl alcohol-water, volume ratio 1.5: 1.5: 5.
5. the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 2 is characterized in that: saussurea involucrata flavonoids dicyandiamide solution is chloroform-methyl alcohol-water, volume ratio 9: 12: 8.
6. the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 2 is characterized in that: earthworm leachate dicyandiamide solution is chloroform-methyl alcohol-water, volume ratio 4: 3: 2.
7. the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 1 is characterized in that: calculate the method error of high speed adverse current chromatogram digitizing dactylogram, formulate the digitizing dactylogram of each batch medicinal material and be made up of following 4 steps;
(1) choose with reference to the peak:
With reference to the peak method, must meet following condition: separate well with adjacent chromatographic peak, peak position is placed in the middle in adopting; Be respectively to wait to reflect the total chromatographic peak of sample; It is the important indicator composition of this kind, if analysis time is long and the peak number order is more, can select twoly with reference to the peak, promptly in the first half chromatographic peak, select and mark the peak one in, in the latter half chromatographic peak, select mark peak in again, form two relative retention value series foundation as a comparison.Relative retention time is retention time and interior recently the quantizing of using chromatographic peak with reference to the peak retention time; Relative peak area adopts with reference to the p-ratio method: as benchmark, making comparisons in the peak area of other each chromatographic peaks and it, obtains a series of area ratio thus with interior peak area with reference to the peak;
(2) foundation of method error
At first will determine the reappearance of method, be similar to and make typical curve, same batch sample is carried out replica test, determine the error of method, the frequency n of revision test should be determined according to mathematical statistics method; Method error comprises as the relative deviation of the relative retention time of qualitative foundation and relative standard deviation, and as the relative deviation and the relative standard deviation of the relative peak area of quantitative basis; If in the method error scope, this group peak of decidable is same peak (component); Then can be considered different component if exceed method error;
(3) make up HSCCC digital finger-print spectrum
Digitizing dactylogram (table 4) is to be based upon on the experimental result basis of chromatographic fingerprinting, when making up the digitizing dactylogram, at first selected with reference to peak i, generally follow following principle: the one, choose the stronger peak in whole chromatogram centre position, the 2nd, better with the degree of separation at two peaks, front and back, the 3rd, this peak is known compound as far as possible, and is important effective constituent;
The calculating of relative retention time is that the retention time with peak i is 1 in the table 4, and with the retention time at other peak by comparison, so the retention time at other peaks all is greater than or less than 1, and mean value is meant the relative retention time t at correspondence peak in the table 4
RMean value, relative retention time t
RRelative standard deviation and relative standard deviation calculate according to following formula;
Relative peak area A adopts the normalized area value in the table 4, promptly the area with reference peak i is 1, with other chromatographic peak areas by comparison, obtain a series of area ratio, variance rate for the same peak of different batches sample is calculated, if determining a certain batch is standard, can compare calculating with it, all not settling the standard as if all batches can be more definite with mean value;
Characteristic fingerprint peak variance rate=| A
I Mark-A
I Mirror|/A
I Mark
The component of relative retention time in the method error scope is placed on a window, promptly thinks to be same component in the method error scope, what exceed thinks different component;
Variance rate is meant each fingerprint peaks and its respective standard fingerprint peaks difference degree between the two.The total variances rate then is the variance rate summation with each fingerprint peaks in the sample.The total variances rate is big more, and expression relatively both sides differs big more, has concluded the comprehensive parameter of qualitative evaluation and quantificational description two aspect information simultaneously;
Characteristic fingerprint peak variance rate=| A
I Mark-A
I Mirror|/A
I Mark
(4) characteristic fingerprint
The general character of a class natural drug sample has been represented at total peak, it also is the characteristic of this kind, be and other kind difference parts, thereby constituted the peculiar fingerprint of this biosome, the peak group that characteristic fingerprint is made up of a series of characteristic fingerprint peak, characteristic fingerprint can be used to contain the discriminating between the different natural drugs of identical effective constituent, being mainly used in kind differentiates, the value difference of fingerprint peaks has reflected the difference of same breed organism, the different place of production, Different Harvesting Time quality, thereby can be used as the Quality Control foundation of raw material and patent medicine again;
The characteristic fingerprint peak must be the common peak of same kind sample, and the peak that has been identified as this medicinal material (medicine materical crude slice) effective constituent or special composition.The characteristic fingerprint peak should account for more than 60% of chromatographic peak sum.The characteristic fingerprint curve is made of characteristic fingerprint and the corresponding area normalization value of each fingerprint peaks, and the situation of certain natural drug characteristic fingerprint is described in the mode of curve map; It is a kind of audio-visual picture, simultaneously the quality and quantity at each characteristic fingerprint peak is shown with the form of curve, and the characteristic fingerprint peak of can will wait to reflect in the same coordinate system system medicine materical crude slice and respective standard shows simultaneously, can be clear that both similarity degrees.
8. the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 1 is characterized in that
The method error of the HSCCC digitizing dactylogram of salvia piece
Method error comprises two aspects: the error range of (1) relative retention time: the relative standard deviation of relative retention time is less than 3%; (2) error range of relative peak area: the relative standard deviation of relative peak area is less than 8%; That is: the RSD of relative retention time＜3% is thought same component; The RSD of relative peak area＜8% thinks that content is identical; The difference degree of each corresponding component in the more different samples;
The HSCCC digitizing dactylogram of salvia piece
There are not non-total peak, peak 1 in the HSCCC dactylogram of red sage root fat-soluble compound
#～12
#All be that these three samples are total, can constitute the characteristic fingerprint of HSCCC, draw the characteristic fingerprint curve with these 12 peaks as the characteristic fingerprint peak.
9. the application of the method for a kind of natural drug high speed counter flow fingerprint spectrum according to claim 1 in the evaluation of the discriminating of natural drug kind, medicinal material differentiation and natural drug composition active component.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103641867A (en) * | 2013-12-05 | 2014-03-19 | 湖南农业大学 | High-speed counter-current chromatography separation method and application of phenolic acid glycoside compounds in traditional Chinese medicine lily |
| CN104031013A (en) * | 2014-06-17 | 2014-09-10 | 浙江工业大学 | Method for preparing salvianolic acid B and rosmarinic acid by adopting high-speed counter-current chromatography separation and purification process |
| WO2020119001A1 (en) * | 2018-12-14 | 2020-06-18 | 香港同盛实业有限公司 | Method for preparing cannabidiol by means of high-speed countercurrent chromatography separation and purification |
| CN112973189A (en) * | 2019-12-13 | 2021-06-18 | 中国科学院大连化学物理研究所 | Method for preparing Chinese medicinal extract absorption component group |
| CN113076812A (en) * | 2021-03-12 | 2021-07-06 | 药都(本溪)一致科技有限公司 | Processing method, system, medium and application of spectrum quantization fingerprint |
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2004
- 2004-12-14 CN CN 200410089500 patent/CN1621832A/en active Pending
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103641867A (en) * | 2013-12-05 | 2014-03-19 | 湖南农业大学 | High-speed counter-current chromatography separation method and application of phenolic acid glycoside compounds in traditional Chinese medicine lily |
| CN103641867B (en) * | 2013-12-05 | 2015-06-24 | 湖南农业大学 | High-speed counter-current chromatography separation method and application of phenolic acid glycoside compounds in traditional Chinese medicine lily |
| CN104031013A (en) * | 2014-06-17 | 2014-09-10 | 浙江工业大学 | Method for preparing salvianolic acid B and rosmarinic acid by adopting high-speed counter-current chromatography separation and purification process |
| CN104031013B (en) * | 2014-06-17 | 2016-08-24 | 浙江工业大学 | A kind of utilize the isolated and purified method preparing salvianolic acid B and rosmarinic acid of high speed adverse current chromatogram |
| WO2020119001A1 (en) * | 2018-12-14 | 2020-06-18 | 香港同盛实业有限公司 | Method for preparing cannabidiol by means of high-speed countercurrent chromatography separation and purification |
| US11267775B2 (en) | 2018-12-14 | 2022-03-08 | Techson Industry Company Limited | Method for preparing cannabidiol by separation and purification using high-speed countercurrent chromatography |
| CN112973189A (en) * | 2019-12-13 | 2021-06-18 | 中国科学院大连化学物理研究所 | Method for preparing Chinese medicinal extract absorption component group |
| CN112973189B (en) * | 2019-12-13 | 2021-12-14 | 中国科学院大连化学物理研究所 | Method for preparing Chinese medicinal extract absorption component group |
| CN113076812A (en) * | 2021-03-12 | 2021-07-06 | 药都(本溪)一致科技有限公司 | Processing method, system, medium and application of spectrum quantization fingerprint |
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