CN101606970A - The method of quality control of radix scutellariae medicinal materials - Google Patents
The method of quality control of radix scutellariae medicinal materials Download PDFInfo
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Abstract
A kind of method of quality control of radix scutellariae medicinal materials is got the standard radix scutellariae medicinal materials, obtains its nuclear magnetic resoance spectrum, and utilizes the characteristic chemical constituent of Radix Scutellariae, points out out the characteristic peak of the characteristic peak of Radix Scutellariae active ingredient as Radix Scutellariae; Get Radix Scutellariae sample to be measured, adopt with the identical method of preparation method, test condition, method of testing of standard radix scutellariae medicinal materials and measure, obtain finger printing, the NMR fingerprint of radix scutellariae medicinal materials to be measured and the finger printing of standard radix scutellariae medicinal materials are compared.According to the difference at the characteristic resonances peak of the finger printing of radix scutellariae medicinal materials to be measured and standard radix scutellariae medicinal materials and the number of common characteristic formant, radix scutellariae medicinal materials can be divided into high-class product N>20, Grade A 18≤N≤20, goods of inferior quality 16≤N<18, certified products 14≤N<16, substandard product N<14.
Description
Technical field
The present invention relates to the method for quality control of Chinese medicine, specifically the method for quality control of radix scutellariae medicinal materials the invention still further relates to the NMR fingerprint of the resulting radix scutellariae medicinal materials of method thus.
Background technology
At present, to the evaluation of radix scutellariae medicinal materials, choose one, two active component or the index components of Radix Scutellariae often and carry out assay, and how much judge quality with its content.If only judge the inherent quality of Radix Scutellariae with the active component of one, two Radix Scutellariae, have certain one-sidedness, said nothing of the index components of no drug effect.Control the effect of radix scutellariae medicinal materials, just can not must be controlled its material group integral body, therefore need to characterize effectively on the whole Chinese medicine quality only at one, two chemical constituents.The material base of Chinese medicine is a chemical constituent, and chemical constituent method by experiment can be characterized by finger printing, and the respective substance basis can be represented in the peak of collection of illustrative plates, if material base composition faciation together, the finger printing that is showed should be consistent.At present, finger printing has become the effective means of internationally recognized control Chinese medicine or natural drug quality.Leaf just very waits (application for a patent for invention number 200610014203.1) people to propose a kind of method of quality control of codonopsis pilosula, employing be the quality of differentiating and estimate medical material by the difference of medical material reference substance liquid-phase chromatograph finger print atlas and medical material liquid-phase chromatograph finger print atlas to be measured relatively.The feasibility of finger printing discriminating Chinese crude drug quality and reliability are based on the detection method that obtains finger printing.The advantage of liquid chromatography detecting method is: liquid chromatograph has good objectivity and The better resolution, disposable Instrument purchase investment is less relatively, but this method belongs to deflection selectivity detection method is arranged, need have the property of intervention to be unfavorable in body and original position analysis sample with destructiveness; Need carry out comparatively complicated preparation and flux is limited to sample; Qualitative (structure is determined) to unknown material has suitable difficulty; The repeatability of method is general.The shortcoming of liquid chromatography detecting method is: there are certain problem in stability before and after stratographic resolution and the chromatographic column sample introduction or repeatability and the qualitative aspects such as (determining structure) of unknown material, all need to be resolved hurrily.In order to overcome the problems referred to above of liquid chromatograph, need to utilize more advanced analysis test method that more objective and accurate Chinese medicine fingerprint is provided.Utilize the NMR fingerprint control Chinese medicine quality of Chinese medicine under this background, to propose just.The advance of Chinese medicine NMR fingerprint and reliability are based on the advantage of nuclear magnetic resonance spectroscopy method of testing.The advantage of magnetic resonance spectroscopy comprises: good objectivity and repeatability, thereby be convenient to the exchange and the comparison of data between the different experiments chamber; Sample does not need loaded down with trivial details processing, can experimentize near under the physiological condition; Have non-invasively, do not destroy structures of samples and character, thereby be convenient to the detection of dynamic of live body, original position; The response coefficient of each active substance is identical in the Chinese medicine extract, therefore can carry out disposable synchronously, do not have the detection of deflection and have good in situ quantitation effect, therefore the application of nuclear magnetic resonance, NMR in the research of complex systems such as Chinese medicine at present is extensive day by day.Simultaneously NMR fingerprint can provide the numerous structural information of Chinese medicine feature extraction thing rapidly reliably, have simple to operate, reliable results, cost is low, advantages such as the many and favorable reproducibility of quantity of information.Therefore, there is strict corresponding relationship in the Chinese medicine feature extraction thing for obtaining under the identical separation program between the NMR fingerprint of Chinese medicine and the herbal species, can be used as the standard of different Chinese medicines of accurate discriminating and the true and false quality of identification Chinese medicine own.
At present to the research of botanical herbs finger printing mainly concentrate on proton nmr spectra (
1HNMR) finger printing aspect, this mainly is because of the required sample size of finger printing also seldom, sweep time is short, obtains easily, so be widely used; But because
1HNMR composes narrow distribution range, therefore causes
1HNMR spectrum fused peaks showed increased forms Bao Feng easily, thereby makes the characteristic signal be difficult to distinguish and point out.In order to overcome above shortcoming, just need to obtain its 13C-NMR (
13CNMR) finger printing, this mainly be because
13CNMR spectrum has the chemical shift range of broad, can distinguish the different carbon atoms that minute differences is structurally arranged in the molecule, and can observe not with proton directly links to each other and contain the carbon functional group, than
1HNMR can provide the structural information of more relevant carbon skeletons, simultaneously
13The probability of CNMR spectrum signal peak overlapping is less, so
13The characteristic signal ratio of CNMR
1HNMR is easier to be distinguished and points out, thereby has higher characteristic.
Summary of the invention
The objective of the invention is by to Radix Scutellariae extract
13A kind of method of Radix Scutellariae quality control is found out in the research of CNMR finger printing, sets up standard radix scutellariae medicinal materials finger printing, and as one of index of quality control and real and fake discrimination, the present invention also provides Radix Scutellariae simultaneously with the finger printing of Radix Scutellariae
13The CNMR standard finger-print.
The present invention implements a kind of method of quality control of radix scutellariae medicinal materials through the following steps, it is characterized in that comprising the steps: that (1) use superconduction pulse Fourier transform NMR spectrometer to set up the 13C-NMR finger printing of standard radix scutellariae medicinal materials and radix scutellariae medicinal materials to be measured with the carbon-13 nuclear magnetic resonance spectrum method; (2) according to the difference at the characteristic resonances peak in radix scutellariae medicinal materials to be measured and the standard radix scutellariae medicinal materials finger printing and the quantity of common characteristic formant, radix scutellariae medicinal materials to be measured can be divided into different qualities.
The described finger printing of setting up the standard radix scutellariae medicinal materials with the carbon-13 nuclear magnetic resonance spectrum method specifically comprises the steps: the preparation of (1) standard radix scutellariae medicinal materials feature extraction thing: get standard radix scutellariae medicinal materials 5g, add 95% ethanol 50mL, reflux is 2 times in water-bath, each 1 hour, the leaching extracting solution, with solvent evaporates, it is standby to obtain standard Radix Scutellariae medical material feature extraction thing;
(2) mensuration of standard radix scutellariae medicinal materials finger printing: precision takes by weighing exsiccant standard Radix Scutellariae medical material feature extraction thing 10.0mg in the nuclear-magnetism test tube, and precision is measured 0.50mL deuterated dimethyl sulfoxide (DMSO-d
6) in the nuclear-magnetism test tube, shake up, measure 25 ℃ of temperature, utilize superconduction pulse Fourier transform NMR spectrometer to obtain the 13C-NMR finger printing of standard radix scutellariae medicinal materials.
The described 13C-NMR finger printing of setting up radix scutellariae medicinal materials to be measured with the carbon-13 nuclear magnetic resonance spectrum method specifically comprises the steps: the preparation of (1) radix scutellariae medicinal materials feature extraction to be measured thing: get radix scutellariae medicinal materials 5g to be measured, add 95% ethanol 50mL, reflux is 2 times in water-bath, each 1 hour, the leaching extracting solution, with solvent evaporates, it is standby to obtain radix scutellariae medicinal materials feature extraction thing to be measured;
(2) mensuration of radix scutellariae medicinal materials finger printing to be measured: precision takes by weighing exsiccant radix scutellariae medicinal materials feature extraction thing 10.0mg to be measured in the nuclear-magnetism test tube, and precision is measured 0.50mLDMSO-d
6In the nuclear-magnetism test tube, shake up, measure 25 ℃ of temperature, utilize superconduction pulse Fourier transform NMR spectrometer to obtain the carbon-13 nmr of radix scutellariae medicinal materials to be measured.
Definite method of common characteristic formant, the common characteristic formant is that the formant of the effective ingredient that contains according to Radix Scutellariae is determined, the characteristic effective ingredient that Radix Scutellariae contains comprises baicalin, oroxylin A, wogonin and baicalin, determines the common characteristic peak of radix scutellariae medicinal materials according to the feature of the feature nuclear magnetic resonance peak of four kinds of effective ingredient in the Radix Scutellariae and finger printing thereof.
Definite method of common characteristic formant number N; There is formant in formant in the finger printing in the scope of characteristic resonances peak left and right sides 0.5ppm, think that promptly this common characteristic formant exists; Radix scutellariae medicinal materials can be divided into the medical material of different qualities, high-class product N>20, Grade A 18≤N≤20, goods of inferior quality 16≤N<18 according to common characteristic formant number N), certified products 14≤N<16, substandard product N<14.
The 13CNMR data of four kinds of characteristic effective ingredient in the table .1 Radix Scutellariae extract
??NO | Baicalin | Oroxylin A, | Wogonin | Baicalin |
??1 | ??182.5 | ??182.2 | ??182.0 | ??182.0 |
??2 | ??163.4 | ??163.2 | ??163.01 | ??163.1 |
??3 | ??153.0 | ??156.5 | ??156.3 | ??151.3 |
??4 | ??152.2 | ??152.3 | ??149.7 | ??148.7 |
??5 | ??147.0 | ??152.6 | ??156.1 | ??146.1 |
??6 | ??132.1 | ??132.2 | ??132.0 | ??131.3 |
??7 | ??131.8 | ??130.8 | ??130.7 | ??103.5 |
??8 | ??129.5 | ??129.2 | ??129.2 | ??128.5 |
??9 | ??129.1 | ??130.2 | ??99.0 | ??128.7 |
??10 | ??126.7 | ??126.4 | ??126.2 | ??125.9 |
??11 | ??105.0 | ??105.1 | ??105.2 | ??105.5 |
??12 | ??104.7 | ??104.6 | ??103.6 | ??104.2 |
??13 | ??94.6 | ??94.5 | ??127.7 | ??93.9 |
??14 | ??60.7 | ??61.0 |
The chemical shift at the total peak of table .2 Radix Scutellariae extract
Table .3 is for chemical shift and the formant number of surveying the total peak of Radix Scutellariae extract
Sequence number | The total formant position/ppm of the feature of contrast Radix Scutellariae | In generation, surveyed the formant position/ppm of Radix Scutellariae | Whether be the characteristic resonances peak |
??1 | ??182 | ??182.2 | Be |
??2 | ??163 | ??163.4 | Be |
??3 | ??157 | ??157.4 | Be |
??4 | ??154 | ??154.5 | Be |
??5 | ??153 | ??153.2 | Be |
??6 | ??152 | ??152.1 | Be |
??7 | ??151 | ??151.5 | Be |
??8 | ??150 | ??150.3 | Be |
??9 | ??149 | ??149.3 | Be |
??10 | ??146 | ??146.7 | Be |
??11 | ??132 | ??132.9 | Be |
??12 | ??131 | ??131.5 | Be |
??13 | ??130 | ??130.5 | Be |
??14 | ??129 | ??129.2 | Be |
??15 | ??128 | ??128.1 | Be |
??16 | ??126 | ??126.1 | Be |
??17 | ??105 | ??105.0 | Be |
??18 | ??103 | ??103.2 | Be |
??19 | ??99 | ??99.4 | Be |
??20 | ??95 | ??95.0 | Be |
??21 | ??93 | ??93.5 | Be |
??22 | ??61 | ??61.1 | Be |
The invention has the beneficial effects as follows:
(1) the Radix Scutellariae NMR finger printing that obtains with Radix Scutellariae extract can be represented most of active ingredient of radix scutellariae medicinal materials, can characterize the quality of radix scutellariae medicinal materials effectively.
(2) do as a wholely to treat with the finger printing of Radix Scutellariae extract, pay attention to the fingerprint characteristic peak and the mutual relation of various chemical constituents, pay attention to the whole facial feature of medical material, both avoided judging the one-sidedness of Radix Scutellariae total quality, and reduced artificial subjective judgment again and cause the probability of error because of only measuring one, two index chemical constituent.The present invention provides new reference standard for quality complete, that accurately estimate Radix Scutellariae, will contribute for improving Radix Scutellariae quality and curative effect.
(3) advantage of the present invention is based on the characteristics of magnetic resonance detection method, nuclear magnetic resonance map has unicity, comprehensive, quantitatively property and the easily characteristics of the property distinguished, the advantage that magnetic resonance detection can provide the multidimensional of reflection molecular characterization to cease characteristic signal more, make the NMR fingerprint authentication technique can provide the number of characteristics signal from many aspects, multi-angle is described the structures of samples feature, thereby makes Chinese medicine NMR fingerprint authentication technique really reach repeated height, characteristic is strong, contain much information, the scope of application is wide, identification result is advantage accurately.
Description of drawings
Fig. 1 is the finger printing of radix scutellariae medicinal materials to be measured and standard radix scutellariae medicinal materials.
The specific embodiment
Embodiment 1: the method that a kind of radix scutellariae medicinal materials finger printing is set up, and it is to take following steps:
(1) foundation of standard radix scutellariae medicinal materials finger printing
The preparation of standard radix scutellariae medicinal materials feature extraction thing:
Get standard radix scutellariae medicinal materials 5g, add 95% ethanol 50mL, reflux is 2 times in water-bath, and each 1 hour, the leaching extracting solution, with solvent evaporates, residue is standby.
The mensuration of standard radix scutellariae medicinal materials finger printing:
Precision takes by weighing 4 hours feature extraction thing 10.0mg of 60 ℃ of dryings in the nuclear-magnetism test tube, and precision is measured 0.5mLDMSO-d
6In the nuclear-magnetism test tube, shake up usefulness to be tested.Condition determination: measure 25 ℃ of temperature, scanning times is 8000 times, DMSO-d
6Make solvent, δ 39.49 residual solvent peaks are interior mark.
(2) foundation of radix scutellariae medicinal materials finger printing to be measured: get radix scutellariae medicinal materials to be measured, measure the finger printing of this Radix Scutellariae medical material to be measured according to Step By Condition described in above-mentioned (1).
(3) in described generation, surveyed the radix scutellariae medicinal materials finger printing and described standard radix scutellariae medicinal materials finger printing compares, discern the common formant that both have and the quantity of common characteristic formant, determine the quality of product quality.
Embodiment 2:(1) standard radix scutellariae medicinal materials sample characteristic extract preparation: get standard radix scutellariae medicinal materials sample 5g, add 95% ethanol 50mL, reflux is 2 times in water-bath, and each 1 hour, the leaching extracting solution, with solvent evaporates, residue is standby.
(2) standard radix scutellariae medicinal materials feature extraction thing preparation: get for test agent 5g, add 95% ethanol 50mL, reflux is 2 times in water-bath, and each 1 hour, the leaching extracting solution, with solvent evaporates, residue is standby.
(3) assay method: precision takes by weighing 4 hours various feature extraction thing 20.0mg of 60 ℃ of dryings in the nuclear-magnetism test tube, and precision is measured 0.5mLDMSO-d
6In the nuclear-magnetism test tube, shake up usefulness to be tested.Condition determination: measure 25 ℃ of temperature, scanning times is 8000 times, DMSO-d
6Make solvent, δ 39.49 residual solvent peaks are interior mark, obtain standard radix scutellariae medicinal materials finger printing.
(4) measure the finger printing of radix scutellariae medicinal materials to be measured according to above-mentioned described Step By Condition.
(5) set up standard radix scutellariae medicinal materials finger printing, the testing sample quality is judged in Radix Scutellariae sample finger printing to be measured and the contrast of standard radix scutellariae medicinal materials finger printing.Described radix scutellariae medicinal materials finger printing to be measured and described standard radix scutellariae medicinal materials finger printing are compared, and discerning the common formant that both have and the quantity of common characteristic formant is 22, and the quality of determining product is an imperial.
The finger printing of radix scutellariae medicinal materials finger printing to be measured and standard radix scutellariae medicinal materials as shown in Figure 1, the formant data as the table .3 shown in.
Claims (5)
1, a kind of method of quality control of radix scutellariae medicinal materials is characterized in that comprising the steps:
(1) use superconduction pulse Fourier transform NMR spectrometer to set up the 13C-NMR finger printing of standard radix scutellariae medicinal materials and radix scutellariae medicinal materials to be measured with the carbon-13 nuclear magnetic resonance spectrum method; (2) according to the difference at the characteristic resonances peak in radix scutellariae medicinal materials to be measured and the standard radix scutellariae medicinal materials finger printing and the quantity of common characteristic formant, radix scutellariae medicinal materials to be measured can be divided into different qualities.
2, the method for quality control of radix scutellariae medicinal materials according to claim 1, it is characterized in that the described finger printing of setting up the standard radix scutellariae medicinal materials with the carbon-13 nuclear magnetic resonance spectrum method specifically comprises the steps: the preparation of (1) standard radix scutellariae medicinal materials feature extraction thing: get standard radix scutellariae medicinal materials 5g, add 95% ethanol 50mL, reflux is 2 times in water-bath, each 1 hour, the leaching extracting solution, with solvent evaporates, it is standby to obtain standard Radix Scutellariae medical material feature extraction thing;
(2) mensuration of standard radix scutellariae medicinal materials finger printing: precision takes by weighing exsiccant standard Radix Scutellariae medical material feature extraction thing 10.0mg in the nuclear-magnetism test tube, and precision is measured 0.50mL DMSO-d
6In the nuclear-magnetism test tube, shake up, measure 25 ℃ of temperature, utilize superconduction pulse Fourier transform NMR spectrometer to obtain the 13C-NMR finger printing of standard radix scutellariae medicinal materials.
3, the method for quality control of radix scutellariae medicinal materials according to claim 1, it is characterized in that the described 13C-NMR finger printing of setting up radix scutellariae medicinal materials to be measured with the carbon-13 nuclear magnetic resonance spectrum method specifically comprises the steps: the preparation of (1) radix scutellariae medicinal materials feature extraction to be measured thing: get radix scutellariae medicinal materials 5g to be measured, add 95% ethanol 50mL, reflux is 2 times in water-bath, each 1 hour, the leaching extracting solution, with solvent evaporates, it is standby to obtain radix scutellariae medicinal materials feature extraction thing to be measured;
(2) mensuration of radix scutellariae medicinal materials finger printing to be measured: precision takes by weighing exsiccant radix scutellariae medicinal materials feature extraction thing 10.0mg to be measured in the nuclear-magnetism test tube, and precision is measured 0.50mL DMSO-d
6In the nuclear-magnetism test tube, shake up, measure 25 ℃ of temperature, utilize superconduction pulse Fourier transform NMR spectrometer to obtain the carbon-13 nmr of radix scutellariae medicinal materials to be measured.
4, the method for quality control of radix scutellariae medicinal materials according to claim 1, the definite method that it is characterized in that the common characteristic formant, the common characteristic formant is that the formant of the effective ingredient that contains according to Radix Scutellariae is determined, the characteristic effective ingredient that Radix Scutellariae contains comprises baicalin, oroxylin A, wogonin and baicalin, determines the common characteristic peak of radix scutellariae medicinal materials according to the feature of the feature nuclear magnetic resonance peak of four kinds of effective ingredient in the Radix Scutellariae and finger printing thereof.
5, the method for quality control of radix scutellariae medicinal materials according to claim 4 is characterized in that definite method of common characteristic formant number N; There is formant in formant in the finger printing in the scope of characteristic resonances peak left and right sides 0.5ppm, think that promptly this common characteristic formant exists; Radix scutellariae medicinal materials can be divided into the medical material of different qualities, high-class product N>20, Grade A 18≤N≤20, goods of inferior quality 16≤N<18, certified products 14≤N<16, substandard product N<14 according to common characteristic formant number N.
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