CN103808749B - A kind of method differentiating capsule of weeping forsythia spin-off - Google Patents

A kind of method differentiating capsule of weeping forsythia spin-off Download PDF

Info

Publication number
CN103808749B
CN103808749B CN201410044108.0A CN201410044108A CN103808749B CN 103808749 B CN103808749 B CN 103808749B CN 201410044108 A CN201410044108 A CN 201410044108A CN 103808749 B CN103808749 B CN 103808749B
Authority
CN
China
Prior art keywords
capsule
weeping forsythia
active component
benzyl carbinol
lignanoid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201410044108.0A
Other languages
Chinese (zh)
Other versions
CN103808749A (en
Inventor
董建军
张海艳
赵天增
郭唯
李晓
陈玲
范毅
常霞
***
魏悦
李倩
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Wang Shuncai
Original Assignee
Henan Kegao Vegetable Natural Product Development Engineering Technology Co ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Henan Kegao Vegetable Natural Product Development Engineering Technology Co ltd filed Critical Henan Kegao Vegetable Natural Product Development Engineering Technology Co ltd
Priority to CN201410044108.0A priority Critical patent/CN103808749B/en
Publication of CN103808749A publication Critical patent/CN103808749A/en
Application granted granted Critical
Publication of CN103808749B publication Critical patent/CN103808749B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Landscapes

  • Medicines Containing Plant Substances (AREA)

Abstract

The present invention relates to a kind of method differentiating capsule of weeping forsythia spin-off, comprising: 1) get capsule of weeping forsythia spin-off directly as the capsule of weeping forsythia lignanoid containing active component group and benzyl carbinol glycosides feature extraction thing; 2) carry out IGD carbon-13 nmr spectra finger-print to described capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing to detect, obtain several active component characteristic peak peak intensities in feature extraction thing according to finger-print; And determine the characteristic peak peak intensity of described each active component respective standard with reference to product by same way; 3) absolute content of described standard with reference to product is obtained by quantitative test means; 4) utilize the ratio of described characteristic peak peak intensity and described absolute content, calculate the content of each active component in capsule of weeping forsythia spin-off and the content of active component group.The present invention can to reflect in capsule of weeping forsythia spin-off containing which lignanoids and benzyl carbinol glycosides compounds and the ratio between them, reaches the object to capsule of weeping forsythia spin-off Quality Identification.

Description

A kind of method differentiating capsule of weeping forsythia spin-off
Technical field
The invention belongs to the discriminating field of natural medicinal plant, particularly, relate to a kind of method differentiating capsule of weeping forsythia spin-off.
Background technology
The capsule of weeping forsythia is that Chinese Clinical commonly uses one of traditional Chinese medicine, for Oleaceae (Oleaceae) Forsythia Vahl (Forsythia) the plant capsule of weeping forsythia (Forsythiasuspensa(Thunb.) Vahl) dry fruit, fruit just ripe gathering when still band is green is called that green grass or young crops sticks up, and gathers to be called and always to stick up during the well-done yellowish of fruit.The capsule of weeping forsythia returns lung, the heart, small intestinl channel, and the property of medicine is slightly cold, bitter, have clearing heat and detoxicating, dispersing swelling and dissipating binds, dispelling wind and heat from the body effect [Chinese Pharmacopoeia Commission. Pharmacopoeia of People's Republic of China version in 2010. China Medical Science Press, 159.].Its active component is mainly lignanoids and benzyl carbinol glycosides compounds, have antiviral, anti-inflammatory, antibacterial, anti-oxidant, fall blood ester, the effect such as to protect the liver [Wang Jinmei, etc. research and development of natural products 2007,19:153.].
Qualitive test certain constituents above-mentioned or its certain active component content of quantitative measurement, become the important method (such as Chinese Pharmacopoeia 2010 editions carries out qualitative and quantitative analysis to lignan component forsythin in capsule of weeping forsythia spin-off and phenylethanoid glycoside Forsythoside) evaluating capsule of weeping forsythia spin-off quality; This quality testing pattern of carrying out qualitative and quantitative analysis for single component effectively can not control the inherent quality of Chinese medical extract, cannot meet current for the objective an urgent demand effectively evaluating and control the capsule of weeping forsythia and the quality of the pharmaceutical preparations thereof.
Finger-print is one of current internationally recognized control Chinese medicine or the most effective mode of natural drug quality.People rely on practical finger-print not only can confirm the true and false of this product, can judge its stability simultaneously.At present; the lignanoid of capsule of weeping forsythia spin-off and the finger-print research work of benzyl carbinol glycosides compounds; concentrate on HPLC finger-print [1. [and Chinese Pharmacopoeia Commission. Pharmacopoeia of People's Republic of China version in 2010. China Medical Science Press; 381. ②Sun states are auspicious; Deng. Chinese patent drug 2007; 29 (2): 161.], capillary electrophoresis fingerprint (HPCE method) [Sun Guoxiang; Deng. chromatogram 2006; 24 (2): 196.], tablets by HPLC-MS (LC-MS) [QuHH; etal.MICROCHEMICALJOURNAL, 2008; 89 (2): 159] etc.Wherein main stream approach HPLC finger-print is owing to affecting by non-chromatographic condition (as chromatographic column internal diameter, length, the Stationary liquid trade mark, carrier fractions, flow rate of mobile phase, the mutually each component ratio of mixed flow, column temperature, sample size, detector sensitivity etc.) comparatively large and need use standard items etc. reason, and repeatability and feasibility all exist many limitation; Though efficient with it, quick, the easy and post of HPCE method not vulnerable to pollution and be better than HPLC method, but still have that reappearance is poor, the range of linearity is narrow and the deficiency such as sensitivity is lower; Single LC-MS analytical approach due to Chinese medicine compound prescription complicacy and lack the present situation of standard items, MS also has the problem such as degree of ionization and matrix interference, be typically not enough to Multiple components is carried out structure-characterized accurately.
IGD carbon-13 nmr spectra coupling (IGD 13cNMRcoupling) fingerprint pattern technology, is also inverted gated decoupling carbon-13 nmr spectra coupling fingerprint pattern technology, this technology be study proton nmr spectra for many years ( 1hNMR) fingerprint pattern technology [Zhao Tianzeng, etc. 1hNMR fingerprint technique plant identification Chinese medicine, Chinese herbal medicine 2000,31 (11): 868-870] basis is combined other technologies (such as current most widely used efficient liquid phase (HPLC) fingerprint pattern technology [Xie Peishan etc., chromatographic fingerprints of Chinese materia medica, People's Health Publisher, 2005] a kind of comprehensive fingerprint pattern technology of non-single means newly) proposed.So far, for capsule of weeping forsythia spin-off and Related product, and have no the disclosure carrying out report and the technology contents differentiated with IGD carbon-13 nmr spectra coupling fingerprint pattern technology.
The quality of capsule of weeping forsythia spin-off does not lie in certain single component, and its curative effect is the result that multiple composition works in coordination with proportioning.Therefore, foundation can not only control capsule of weeping forsythia spin-off active component, and the IGD carbon-13 nmr spectra coupling finger-print of the ratio that can control these active components is imperative.The research and apply of capsule of weeping forsythia spin-off carbon-13 nmr spectra coupling finger-print, not only can solve the difficult problem that China's capsule of weeping forsythia spin-off is evaluated, also for strengthening systematization and the standardization of the inherent composition Study of capsule of weeping forsythia spin-off, the guarantee providing science in line with international standards is realized.Along with this technology applying in other Chinese medical extracts, the great scientific value of this technology will be increasingly outstanding.
Summary of the invention
In order to solve the problem of prior art, the object of this invention is to provide a kind of method differentiating capsule of weeping forsythia spin-off.
To achieve these goals, the method for discriminating capsule of weeping forsythia spin-off provided by the invention, comprises the following steps:
1) capsule of weeping forsythia spin-off is got directly as the capsule of weeping forsythia lignanoid containing active component group (capsule of weeping forsythia lignanoid and benzyl carbinol glycosides) and benzyl carbinol glycosides feature extraction thing;
2) carry out IGD carbon-13 nmr spectra finger-print to described capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing to detect, obtain several active component characteristic peak peak intensities in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing according to finger-print; And determine the characteristic peak peak intensity of described each active component respective standard with reference to product by same way (IGD carbon-13 nmr spectra finger-print);
3) absolute content of described standard with reference to product is obtained by quantitative test means;
4) ratio of described characteristic peak peak intensity (the characteristic peak peak intensities of each active component characteristic peak peak intensity and respective standard reference product) and the absolute content of described standard reference product is utilized, calculate the content of each active component and the total content of this active component, the i.e. content of active component group in capsule of weeping forsythia spin-off.
Wherein, adopt there is the extraction process that the obtains clear IGD carbon-13 nmr spectra finger-print extracting mode as described capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing.
Wherein, step 2) in, carry out IGD carbon-13 nmr spectra finger-print to capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing and detect, the solvent dissolving capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing is deuterated methanol (CD 3or deuterated dimethyl sulfoxide (DMSO-d OD) 6), preferred deuterated dimethyl sulfoxide; The mass volume ratio of capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing and coordinative solvent is 55:1 ~ 65:1(mg:mL), preferred 60:1.
Wherein, step 2) in, in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing, the characteristic peak of lignanoids active component is C-7 absorption peak, and its chemical shift is δ c83.0 ~ 88.0.
Preferably, select C-7 ' distinguish the forsythin in lignanoids active component further as characteristic peak and show rosin element-4-β-D-Glucose glycosides, their chemical shift is δ c81.0 ~ 82.0.
In capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing, the characteristic peak of benzyl carbinol glycosides active component is C-9 ' absorption peak, and its chemical shift is δ c165.0 ~ 170.0.
Preferably, select C-1 or C-6 to distinguish forsythiaside A in benzyl carbinol glycosides active component and Forsythoside H further as characteristic peak, their chemical shift is respectively δ c119.0 ~ 120.0 and δ c129.0 ~ 130.0.
Wherein, step 2) in, according to size and the position of characteristic peak peak intensity, several active components in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing are sorted.
Further, step 2) in, according to size and the position of characteristic peak peak intensity, the active component in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing: forsythin, table rosin element-4-β-D-Glucose glycosides, loose ester element-β-D-Glucose glycosides, Forsythingenin, "-O-glucoside, forsythiaside A, Forsythoside H, Forsythoside I, Forsythoside F's 1-hydroxyl-rosin element 4 sort.
Wherein, step 2) in, described peak intensity can adopt peak height method, area integral method or gravimetric method to calculate.
Wherein, in step 3), described quantitative test means are: efficient liquid phase (HPLC) method.
Further, when differentiating capsule of weeping forsythia lignanoids active component, the condition of described high-efficient liquid phase technique comprises:
Mobile phase is acetonitrile: water=(20 ~ 30): (70 ~ 80), preferred 25:75; Determined wavelength is 277nm.
Further, when differentiating benzyl carbinol glycosides active component, the condition of described high-efficient liquid phase technique comprises:
Mobile phase is acetonitrile: glacial acetic acid solution=(10 ~ 20): (80 ~ 90), preferred 15:85; Determined wavelength is 330nm.
Wherein, in step 3), described standard refers to reference to the absolute content of product: the standard measured by quantitative test means is with reference to the mass percentage of product.
Further, in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing, the standard of lignanoids active component and benzyl carbinol glycosides active component is forsythin or forsythiaside A with reference to product.
Wherein, in step 4), the coupling formula calculating the content of each active component is:
W n = W 1 M n h n M 1 h 1 ; Wherein:
W 1for standard corresponding to a certain active component in the capsule of weeping forsythia spin-off that step 3) quantitative test means measure is with reference to the absolute content of product;
M 1for standard corresponding to a certain active component in described capsule of weeping forsythia spin-off is with reference to the carbon number that molecular weight/quantitatively peak is corresponding of product;
H 1for standard corresponding to a certain active component in the capsule of weeping forsythia spin-off by IGD carbon-13 nmr spectra determining fingerprint pattern is with reference to the characteristic peak peak intensity of product;
W nfor the mass percentage of active component a certain in capsule of weeping forsythia spin-off;
M nfor the carbon number that molecular weight/quantitatively peak is corresponding of active component a certain in capsule of weeping forsythia spin-off;
H nfor the characteristic peak peak intensity of a certain active component in the capsule of weeping forsythia spin-off by IGD carbon-13 nmr spectra determining fingerprint pattern; The total content of this active component is exactly the W of similar each active component nsum, the i.e. content of active component group.
The derivation of above-mentioned formula is:
Active component group described in the inventive method is the active component group in capsule of weeping forsythia spin-off.Described capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing refer to that namely active component group refers to the summation of corresponding same active component containing capsule of weeping forsythia lignanoid and benzyl carbinol glycosides active component in this extract.
Wherein, described capsule of weeping forsythia spin-off comprises: forsythia suspense extraction or capsule of weeping forsythia natural drug.
Forsythia suspense extraction of the present invention or spin-off, comprise each position of capsule of weeping forsythia plant, as extract or the spin-off of root, skin, stem, leaf, flower and fruit etc.
Further, described forsythia suspense extraction can comprise: capsule of weeping forsythia berry extract, capsule of weeping forsythia lignanoid or benzyl carbinol glycosides extract, chromocor extract, triterpenic acid extract, or Folium Forsythiae extract etc.
The calculating of the content of each active component of the present invention and the total content of this active component is by IGD carbon-13 nmr spectra and the coupling of analysis quantitative means by coupling formula.Compared to the prior art, the present invention adopts IGD 13cNMR coupling finger-print has several feature below:
1. stability (repeatability): IGD 13the chemical shift data that CNMR obtains is second after radix point, and explanation property is good, reproducible; The non-chromatographic condition (as chromatographic column internal diameter, length, the Stationary liquid trade mark, carrier fractions, flow rate of mobile phase, the mutually each component ratio of mixed flow, column temperature, sample size, detector sensitivity etc.) of HPLC, GC changes, the retention time data variation obtained is very large, mean the variation of monolithic chromatogram figure, repeatability is bad.
2. globality (comprehensive): IGD 13the corresponding spectrum peak of each the active component carbon in sample is comprised in CNMR finger-print; There is not this relation in HPLC, GC, UV, IR, MS.
3. reliability (unicity): IGD 13it is strict one-to-one relationship from the carbon on different activities composition in sample and different group thereof that CNMR composes peak; There is not this relation in HPLC, GC, UV, IR, MS.
4. feasibility (easily the property distinguished): IGD 13cNMR finger-print regularity is very strong, generally, can belong to each the carbon peak in collection of illustrative plates; HPLC, GC need reference substance; IR not easily resolves; UV quantity of information is few; MS then has the problem such as degree of ionization and matrix interference.
IGD carbon-13 nmr spectra finger-print can only show there is which active component in feature extraction thing, and the quantitative ratio between these active components, and the absolute content of these active components must analyze quantitative means by standard with reference to product and other, then obtained by coupling formula.
The advantage that the present invention has is as follows:
1, four steps of the present invention are an entirety, indispensable; Four steps have respective distinctive feature; If separately or simple combination then can not detect ratio and the content of complicated ingredient in medicinal plant.
As everyone knows, although Chinese medicine, botanical pesticide are time-honored medicines, it is analyzed and detects is a difficult problem always.Therefore, special need a kind of method can various active components in differential plant kind and extract thereof, and it is quantitative, then select best extract component or its composition, to give full play to its effect.In addition, one of obstacle that Chinese medicine, botanical pesticide are gone abroad does not have constituent and quantitative identifying thereof, and the qualitative and quantitative analysis aspect that reason is currently available technology solution differential plant kind and evaluation plant source product quality also exists significant limitation (specifically seeing the content that the application's background technology describes).Fact proved, the gordian technique that IGD carbon-13 nmr spectra coupling fingerprint pattern technology addresses this problem just, this is seeing existing explanation in present specification.
Although 2 traditional extraction techniques are known by those of ordinary skill, people is never had to obtain clear collection of illustrative plates and representative active component group obtains feature extraction thing as standard to select technological parameter.Utilize the method, the identification result finally for extract can be made better, and this is one of difficult point of the present invention, is also one of innovative point of the present invention.
3, IGD 13cNMR coupling finger-print is the mixed spectrum of multiple active component, inevitably causes the crowded of excellent peak one by one, even overlapping.In order to make result of calculation accurate, specific characteristic peak, each active component carbon peak in the active component group that chemical shift difference is larger is selected to be necessary.Because dissimilar compound carbon spectral difference is not very large, the selection of characteristic peak can be ever-changing; Identical type compound carbon spectral difference is not very little, and a lot of peak overlap is serious, deep nuclear magnetic resonance spectrum knowledge need be had to instruct, through in many ways comparing, turning over, could select good characteristic peak; And characteristic peak select bad, have no idea to carry out accurate quantitative analysis to active component group, this also just the present invention need solve problem.According to the feature of lignanoids in the capsule of weeping forsythia and phenylethanoid glycoside, select C-7 to distinguish lignan component as characteristic peak, and select C-7 ' to distinguish this overlapping constituents further as characteristic peak; Select C-9 ' as characteristic peak difference phenylethanoid glycoside, and select C-1 or C-6 to distinguish this overlapping constituents further as characteristic peak.So, need to select different specific characteristic peaks, active component carbon peak to be also the another innovative point of the application according to the feature of different activities composition.
4, because characteristic peak chemical shift difference is very little, in a lot of situation, only after radix point, the 1st potential difference is other, so the sequence of characteristic peak is the key determining main active and ratio thereof, there is no deep nuclear magnetic resonance spectrum knowledge and be separated basis, the active component being difficult to determine that characteristic peak represents and ratio thereof, also just accurate qualitative and quantitative analysis cannot be carried out to active component group, according to the feature of the carbon spectrum nuclear magnetic data of lignanoids in the capsule of weeping forsythia and phenylethanoid glycoside, accurate sequence is carried out to the characteristic peak of two constituents; This is one of difficult point of the present invention, is also one of innovative point of the present invention.
5, coupling calculates the selection that key is choice criteria product and quantitative analysis tech.Analyze quantitative means can select efficient liquid phase, gas chromatography, thin-layered chromatography and weighing method etc., standard with reference to product can be a certain active component as interior mark, also can be additional with reference to product as external standard.According to the feature of lignanoids in the capsule of weeping forsythia and phenylethanoid glycoside, what analyze that quantitative means selects be liquid phase, and standard is forsythin or Forsythoside (can have different choice according to different situations) with reference to product selection.This is one of difficult point of the present invention, is also one of innovative point of the present invention.
The present invention adopts IGD carbon-13 nmr spectra finger-print to differentiate capsule of weeping forsythia spin-off, can to reflect in capsule of weeping forsythia spin-off containing which capsule of weeping forsythia lignanoids and phenylethyl alcohol glycoside and the ratio between them, reach the object to capsule of weeping forsythia spin-off Quality Identification.The range of linearity is wide, highly sensitive, repeatability and feasibility good.Effectively can not only control the inherent quality of capsule of weeping forsythia spin-off, also can meet current for the objective an urgent demand effectively evaluated and control capsule of weeping forsythia quality.All in all, not only can solving the difficult problem that China's capsule of weeping forsythia spin-off is evaluated, also for strengthening systematization and the standardization of the inherent composition Study of capsule of weeping forsythia spin-off, realizing the guarantee providing science in line with international standards.
Accompanying drawing explanation
Fig. 1-a is the IGD carbon-13 nmr spectra finger-print that embodiment 1 makes Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract by oneself.
Fig. 1-b, 1-c and 1-d be embodiment 1 make by oneself Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract IGD carbon-13 nmr spectra Fingerprints peak local widen enlarged drawing.
Fig. 2-a is the IGD carbon-13 nmr spectra finger-print of the commercially available forsythia suspense extraction of embodiment 2.
Fig. 2-b and 2-c is that the local, IGD carbon-13 nmr spectra Fingerprints peak of the commercially available forsythia suspense extraction of embodiment 2 widens enlarged drawing.
Fig. 3-a is the IGD carbon-13 nmr spectra finger-print that embodiment 3 makes Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract first (precipitation method) by oneself.
Fig. 3-b, 3-c and 3-d be embodiment 3 make by oneself Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract first (precipitation method) IGD carbon-13 nmr spectra Fingerprints peak local widen enlarged drawing.
Fig. 4-a is that embodiment 4 makes Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides extract (resin method) IGD carbon-13 nmr spectra finger-print by oneself.
Fig. 4-b, 4-c and 4-d are that embodiment 4 makes Luanchuan, Yuzhou Folium Forsythia lignanoid by oneself and local, benzyl carbinol glycosides extract (resin method) IGD carbon-13 nmr spectra Fingerprints peak widens enlarged drawing.
Fig. 5-a is the IGD carbon-13 nmr spectra finger-print that embodiment 5 makes Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second (resin method) by oneself.
Fig. 5-b, 5-c and 5-d be embodiment 5 make by oneself Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second (resin method) IGD carbon-13 nmr spectra Fingerprints peak local widen enlarged drawing.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
If without particularly pointing out, the material that the present invention uses is the conventional material of this area, and NM method of operating is also the method for this area routine.
In the present invention, the concentration of ethanol is percent by volume.
One, capsule of weeping forsythia spin-off IGD carbon-13 nmr spectra finger-print research
(1) acquisition of capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing (CET)
Get capsule of weeping forsythia spin-off directly as capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing (CET)
(2) the IGD carbon-13 nmr spectra finger-print of capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing detects
Get capsule of weeping forsythia lignanoid and/or benzyl carbinol glycosides feature extraction thing 30mg, be dissolved in 0.5mL deuterated dimethyl sulfoxide (DMSO-d 6) or deuterated methanol in, do IGD carbon-13 nmr spectra finger-print detect, Ji get capsule of weeping forsythia lignanoid and benzyl carbinol glycosides IGD carbon-13 nmr spectra finger-print.
(3) the IGD carbon-13 nmr spectra finger-print of capsule of weeping forsythia lignanoids and benzyl carbinol glycosides feature extraction thing is resolved
1. differentiate
In capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing (CET) IGD carbon-13 nmr spectra finger-print, clearly illustrate the characteristic signal of capsule of weeping forsythia lignanoids and benzyl carbinol glycosides compounds, forsythin, loose ester element-β-D-Glucose glycosides, forsythiaside A, Forsythoside H, Forsythoside I etc. all have corresponding NMR signal.
2. each active component characteristic peak in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing is chosen
Need to select different specific characteristic peaks, active component carbon peak according to the feature of different activities composition.Selection principle is as follows: 1. the specific characteristic peak of similar compound is preferably identical carbon carbon peak, each compound position; 2. between the specific characteristic peak of each compound and other carbon peaks, chemical shift difference is larger; Between the specific characteristic peak of 3. each compound, chemical shift difference is larger; 4. the chemical shift effect difference affecting the specific characteristic peak of each compound itself is larger.
Owing to containing a series of lignanoids and benzyl carbinol glycosides compounds (active component) in capsule of weeping forsythia lignanoids and benzyl carbinol glycosides feature extraction thing, carbon peak intersects more, in order to measure the ratio of each active component, the larger respective peaks of chemical shift difference must be selected as characteristic peak., investigate through reality for this reason, select δ cδ, as specific characteristic peak, lignanoids active component carbon peak, is selected in about 83.0 ~ 88.0 one groups of C-7 tertiary carbon carbon peaks cabout 165.0 ~ 170.0 one groups of C-9 ' carbonyl carbon carbon peaks are as specific characteristic peak, benzyl carbinol glycosides active component carbon peak; Its reason is that Lignanoids compounds C-7 tertiary carbon and benzyl carbinol glycosides compounds C-9 ' carbonyl carbon and other carbon chemical shifts difference are comparatively large, easy to identify; And chemical shift also has certain difference between dissimilar compound and different Compound C-9 ' carbonyl carbon carbon peak of the same type.Preferably, C-7 ' can be selected to distinguish forsythin and the table rosin element-4-β-D-Glucose glycosides of C-7 characteristic peak overlap in lignanoids active component further as characteristic peak, and their chemical shift is δ c81.0 ~ 82.0; C-1 or C-6 can be selected to distinguish forsythiaside A and the Forsythoside H of the C-9 ' characteristic peak overlap in benzyl carbinol glycosides active component further as characteristic peak, and their chemical shift is respectively δ c119.0 ~ 120.0 and δ c129.0 ~ 130.0.
3. standard is with reference to the selection of product
Forsythin is capsule of weeping forsythia medicinal material and extract thereof, and main lignanoids active component in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing, and its C-7 chemical shift is about δ c86.7(DMSO-d 6) do not have overlapping at this with other main lignanoids and phenylethanoid glycoside; Forsythiaside A is main benzyl carbinol glycosides active component in capsule of weeping forsythia medicinal material and extract thereof, and the chemical shift of itself C-1 or C-6 is respectively δ c119.0 ~ 120.0 and δ c129.0 ~ 130.0, do not have overlapping at this with other main lignanoids and phenylethanoid glycoside; Therefore, forsythin or Forsythoside is selected as the standard of 2 active components with reference to product.
(4) HPLC method is adopted to measure the content of forsythin and Forsythoside in capsule of weeping forsythia spin-off
1. forsythin
I) chromatographic condition (with reference to 2010 editions pharmacopeia)
Mobile phase is acetonitrile: water=25:75;
Chromatographic column: C18(250*4.6mm, 5um);
Determined wavelength: 277nm.
Ii) forsythin standard is with reference to the preparation of product solution
Accurately take forsythin 5mg, put in 50mL volumetric flask, with methyl alcohol dissolved dilution to scale, namely obtain forsythin standard after shaking up with reference to product solution.
The preparation of the need testing solution iii) containing capsule of weeping forsythia lignanoids active component
Accurately take test sample: capsule of weeping forsythia spin-off (as forsythia suspense extraction) 200mg is in 100mL volumetric flask, add proper amount of methanol (30-40mL) to dissolve, scale is diluted to, i.e. capsule of weeping forsythia spin-off need testing solution (as forsythia suspense extraction need testing solution) after sonic oscillation.
Iv) determination method
Accurate forsythin standard of drawing is with reference to product solution and need testing solution (i.e. above-mentioned capsule of weeping forsythia spin-off need testing solution) each 20 μ 1 containing capsule of weeping forsythia lignanoids active component respectively, and injection liquid chromatography, measures, obtain Determination of forsythin.
V) in capsule of weeping forsythia spin-off, forsythin absolute content calculates
Forsythin mass concentration in the need testing solution containing lignanoids active component is calculated by following formula:
C X = C R × A X A R
C x: forsythin mass concentration (ug/mL) in the need testing solution containing lignanoids active component;
C r: forsythin standard is with reference to product concentration of polymer solution (ug/mL);
A x: the peak area of the need testing solution containing lignanoids active component;
A r: forsythin standard is with reference to the peak area of product solution.
Vi) forsythin mass percentage in capsule of weeping forsythia spin-off is calculated by following formula
W forsythin(%): forsythin mass percentage in capsule of weeping forsythia spin-off;
C x: the forsythin mass concentration (ug/mL) in the need testing solution containing lignanoids active component;
M: the quality (mg) of the capsule of weeping forsythia spin-off taken.
2. Forsythoside assay
I) chromatographic condition (with reference to 2010 editions pharmacopeia)
Mobile phase is acetonitrile: glacial acetic acid solution=15:85;
Chromatographic column: C18(250*4.6mm, 5um);
Determined wavelength: 330nm.
Ii) Forsythoside standard is with reference to the preparation of product solution
Accurately take Forsythoside 5mg, put in 50mL volumetric flask, with methyl alcohol dissolved dilution to scale, namely obtain Forsythoside standard after shaking up with reference to product solution.
The preparation of the need testing solution iii) containing capsule of weeping forsythia benzyl carbinol glycosides active component
Accurately take test sample: capsule of weeping forsythia spin-off (as forsythia suspense extraction) 200mg is in 100mL volumetric flask, add proper amount of methanol (30-40mL) to dissolve, scale is diluted to, i.e. capsule of weeping forsythia spin-off need testing solution (as forsythia suspense extraction need testing solution) after sonic oscillation.
Iv) determination method
Accurate Forsythoside standard of drawing is with reference to product solution and need testing solution (i.e. above-mentioned capsule of weeping forsythia spin-off need testing solution) each 20 μ 1 containing capsule of weeping forsythia benzyl carbinol glycosides active component respectively, and injection liquid chromatography, measures, obtain Determination of forsythin.
V) in capsule of weeping forsythia spin-off, Forsythoside absolute content calculates
Forsythin mass concentration in the need testing solution containing benzyl carbinol glycosides active component is calculated by following formula:
C X = C R × A X A R
C x: Forsythoside mass concentration (ug/mL) in the need testing solution containing benzyl carbinol glycosides active component;
C r: Forsythoside standard is with reference to product concentration of polymer solution (ug/mL);
A x: the peak area of the need testing solution containing benzyl carbinol glycosides active component;
A r: Forsythoside standard is with reference to the peak area of product solution.
Vi) Forsythoside mass percentage in capsule of weeping forsythia spin-off is calculated by following formula
W forsythoside(%): Forsythoside mass percentage in capsule of weeping forsythia spin-off;
C x: the Forsythoside mass concentration (ug/mL) in the need testing solution containing benzyl carbinol glycosides active component;
M: the quality (mg) of the capsule of weeping forsythia spin-off taken.
(5) by content and total amount, the i.e. content of lignanoids or benzyl carbinol glycosides active component group of lignanoids or each main active of phenylethyl alcohol glycoside in coupling formulae discovery capsule of weeping forsythia spin-off
W n = W 1 M n h n M 1 h 1
W 1: the mass percentage of capsule of weeping forsythia spin-off Plays reference product forsythin or Forsythoside;
M 1: the carbon number that molecular weight/quantitatively peak is corresponding of standard reference product forsythin or Forsythoside;
H 1: by the standard reference product forsythin of IGD carbon-13 nmr spectra determining fingerprint pattern or the characteristic peak peak intensity (peak height) of Forsythoside;
W n: the mass percentage of a certain lignanoids or benzyl carbinol glycosides active component in capsule of weeping forsythia spin-off;
M n: the carbon number that molecular weight/quantitatively peak is corresponding of a certain lignanoids or benzyl carbinol glycosides active component in capsule of weeping forsythia spin-off;
H n: by a certain lignanoids or benzyl carbinol glycosides active component characteristic peak peak intensity (peak height) in the capsule of weeping forsythia spin-off of IGD carbon-13 nmr spectra determining fingerprint pattern.
Two, instrument, reagent and material
Key instrument and equipment
Nuclear magnetic resonance spectrometer BrukerDPX400 type.
Mass spectrometer: WatersMicromass company Q-TofMicroTM type.
Half preparative high-performance liquid chromatographic instrument: Waters600 type.
High performance liquid chromatograph: Agilent1200 type.
2000mL distilling flask, 5000mL distilling flask, spherical condensating tube, 2000mL separating funnel.
DE-52AA Rotary Evaporators: Shanghai Yarong Biochemical Instrument Plant.
DEF-6020 type vacuum drying chamber: the upper grand experimental facilities company limited of Nereid.
Green grass or young crops sticks up, always stick up (Luanchuan In Henan, Yuzhou are plucked from local for 2012), commercially available forsythia suspense extraction (Kang Peiji bio tech ltd, Xi'an), and medicinal material is identified through In Henan Agriculture university professor Zhu Changshan; Forsythin, chemical reference substance, laboratory self-control (through spectroscopic data qualification).
Reagent: chromatographically pure (methyl alcohol, Tianjin Siyou Fine Chemicals Co., Ltd.) and analyzing pure (Tianjin Chemical Reagents Factory No.1).
Three, fundamental research
(1) structure of main active and carbon-13 nmr spectra data in capsule of weeping forsythia spin-off
Forsythin: R 1=Me, R 2=Glc
Forsythingenin: R 1=Me, R 2=H
Table rosin element-4-β-D-Glucose glycosides: R 1=H, R 2=Glc
Show loose ester element-4 '-O-glucoside: R 1=Glc, R 2=H
Pine ester element-β-D-Glucose glycosides: R 1=H, R 2=Glc
1-hydroxyl-rosin element 4 "-O-glucoside
The blue A:R=H of the capsule of weeping forsythia
The blue B:R=Me of the capsule of weeping forsythia
Forsythiaside A
Forsythiaside B
Forsythoside H
Forsythoside F
Forsythoside I
Forsythoside J
N-butoxy Forsythoside
Forsythin (Phillyrin)
13CNMR(100MHz,DMSO-d 6)δ C:135.2(C-1),110.3(C-2),148.8
(C-3),145.8(C-4),115.1(C-5),118.0(C-6),86.5(C-7),53.9
(C-8),70.2(C-9),131.2(C-1′),109.3(C-2′),148.5(C-3′),146.0
(C-4′),111.4(C-5′),117.4(C-6′),81.1(C-7′),49.2(C-8′),68.9
(C-9′),55.4,55.4,55.6(3OMe)，100.0(Glc-1),73.1(Glc-2),76.7
(Glc-3),69.6(Glc-4),76.9(Glc-5),60.6(Glc-6)。
Forsythingenin (Phillygenin)
13CNMR(100MHz,DMSO-d 6)δ C:132.2(C-1),110.2(C-2),147.4*
(C-3),145.9(C-4),115.1(C-5),118.5(C-6),86.9(C-7),53.8
(C-8),70.2(C-9),131.1(C-1′),109.4(C-2′),148.4(C-3′),147.6*
(C-4′),111.5(C-5′),117.5(C-6′),81.3(C-7′),49.3(C-8′),68.7
(C-9′),55.4,55.4,55.5(3OMe)。(* ownership is interchangeable)
Table rosin element-4-β-D-Glucose glycosides (epipinoresinol-4-β-D-glucoside)
13CNMR(100MHz,DMSO-d 6)δ C:135.3(C-1),110.4(C-2),148.9(C-3),145.9(C-4),115.1(C-5),118.1*(C-6),86.6(C-7),54.0(C-8),70.2(C-9),129.6(C-1′),109.7(C-2′),147.3(C-3′),145.2(C-4′),115.1(C-5′),117.9*(C-6′),81.4(C-7′),49.3(C-8′),69.0(C-9′),55.5,55.6(2OMe)，100.1(Glc-1),73.2(Glc-2),76.8**(Glc-3),69.6(Glc-4),77.0*(Glc-5),60.6(Glc-6)。(* ownership is interchangeable)
Pine ester element-β-D-Glucose glycosides [(+)-pinoresinol-β-D-glucoside]
13CNMR(100MHz,DMSO-d 6)δ C:135.2(C-1),115.1(C-2),148.9(C-3),145.9(C-4),114.8(C-5),118.6(C-6),85.1(C-7),53.5(C-8),70.9(C-9),132.1(C-1′),110.4(C-2′),148.9(C-3′),148.9(C-4′),115.1(C-5′),118.6(C-6′),84.8(C-7′),53.5(C-8′),70.9(C-9′),55.6(2OMe)，100.0(Glc-1),73.1(Glc-2),76.9(Glc-3),69.6(Glc-4),76.9(Glc-5),60.6(Glc-6)。(* ownership is interchangeable)
1-hydroxyl-rosin element 4 "-O-glucoside (Hydroxypinoresinol-4 "-O-β-D-glucoside)
13CNMR(100MHz,DMSO-d 6)δ C:135.3(C-1),110.9(C-2),148.9(C-3),145.9(C-4),115.2(C-5),118.4(C-6),85.1(C-7),60.8(C-8),70.3(C-9),128.0(C-1′),112.4(C-2′),146.9(C-3′),146.0(C-4′),114.6(C-5′),120.2(C-6′),87.2(C-7′),91.0(C-8′),74.8(C-9′),55.6,55.8(2OMe)，100.1(Glc-1),73.2(Glc-2),76.8(Glc-3),69.6(Glc-4),77.0(Glc-5),60.6(Glc-6)。(* ownership is interchangeable)
The blue A(forsythialanA of the capsule of weeping forsythia)
13CNMR(75MHz,CDCL 3)δ:132.30(C-1),110.04(C-2),146.84(C-3),114.06(C-4),145.57(C-5),120.08(C-6),83.85(C-7),52.22(C-8),61.35(C-9),129.42(C-1′),110.39(C-2′),146.85(C-3′),150.84(C-4′),114.07(C-5′),123.81(C-6′),197.99(C-7′),49.62(C-8′),70.78(C-9′)，56.07(OCH 3-3)，55.96(OCH 3-3′)。
The blue B(forsythialanB of the capsule of weeping forsythia)
13CNMR(75MHz,CDCL 3)δ:132.32(C-1),108.97(C-2),146.85(C-3),114.04(C-4),145.62(C-5),120.11(C-6),83.88(C-7),52.25(C-8),61.42(C-9),129.79(C-1′),110.67(C-2′),149.27(C-3′),153.71(C-4′),10.15(C-5′),123.16(C-6′),197.96(C-7′),49.66(C-8′),70.80(C-9′)，56.01(OCH 3-3)，56.01(OCH 3-3′)。
Show loose ester element-4 '-O-glucoside (epipinoresinol-4 '-O-glucoside)
13CNMR(100MHz,DMSO-d 6)δ C:132.2*(C-1),110.2(C-2),147.4(C-3),145.9(C-4),114.8(C-5),118.6(C-6),86.9(C-7),53.8(C-8),68.8(C-9),132.3*(C-1′),109.9(C-2′),148.5(C-3′),145.4(C-4′),115.1(C-5′),117.5(C-6′),81.1(C-7′),49.2(C-8′),68.8(C-9′),55.5,55.6(2OMe)，100.0(Glc-1),73.2(Glc-2),76.8(Glc-3),69.6(Glc-4),76.9(Glc-5),60.6(Glc-6)。(* ownership is interchangeable)
Forsythiaside A [ForsythosideA (forsythiaside)]
13CNMR(100Mz,DMSO)δ:129.2(C-1),115.5(C-2),144.9(C-3),143.5(C-4),116.3(C-5),119.5(C-6),35.1(C-7),70.3(C-8),125.4(C-1′),114.8(C-2′),145.6(C-3′),148.6(C-4′),115.8(C-5′),121.4(C-6′),145.6(C-7′),113.6(C-8′),165.9(C-9′),102.9(Glc-1),73.0(Glc-2),73.5(Glc-3),71.0(Glc-4),73.9(Glc-5),66.1(Glc-6),100.5(Rha-1),70.6(Rha-2),70.6(Rha-3),71.9(Rha-4),68.4(Rha-5),17.8(Rha-6)
13CNMR(100Mz,CD 3OD)δ:131.4(C-1),116.4(C-2),146.1(C-3),144.7(C-4),117.2(C-5),121.3(C-6),36.7(C-7),72.4(C-8),127.7(C-1′),115.3(C-2′),146.9(C-3′),149.8(C-4′),116.6(C-5′),123.2(C-6′),147.7(C-7′),114.7(C-8′),168.3(C-9′),104.5(Glc-1),75.2(Glc-2),75.8(Glc-3),72.4(Glc-4),74.8(Glc-5),67.7(Glc-6),102.3(Rha-1),72.3(Rha-2),
72.1(Rha-3),74.0(Rha-4),69.9(Rha-5),18.0(Rha-6)。
Forsythiaside B (ForsythosideB)
13CNMR(125Mz,CD 3OD)δ:131.4(C-1),115.3(C-2),146.0(C-3),144.5(C-4),116.3(C-5),121.3(C-6),36.5(C-7),72.2(C-8),127.6(C-1′),114.7(C-2′),146.7(C-3′),149.6(C-4′),116.5(C-5′),123.2(C-6′),148.0(C-7′),117.1(C-8′),168.2(C-9′),104.1(Glc-1),81.6(Glc-2),76.0(Glc-3),70.4(Glc-4),74.4(Glc-5),68.3(Glc-6),103.0(Rha-1),72.2(Rha-2),72.0(Rha-3),73.7(Rha-4),70.8(Rha-5),18.4(Rha-6)，110.9(APi-1),78.0(APi-2),80.6(APi-3),75.0(APi-4),65.6(APi-5)。
Forsythoside H(ForsythosideH)
13CNMR(100Mz,DMSO)δ:129.1(C-1),115.4(C-2),144.9(C-3),143.5(C-4),116.2(C-5),119.6(C-6),35.1(C-7),69.8(C-8),125.5(C-1′),114.8(C-2′),145.6(C-3′),148.5(C-4′),115.8(C-5′),121.3(C-6′),145.2(C-7′),114.2(C-8′),165.7(C-9′),100.2(Glc-1),73.4(Glc-2),74.1(Glc-3),70.7(Glc-4),75.5(Glc-5),66.6(Glc-6),100.7(Rha-1),70.5(Rha-2),70.3(Rha-3),72.0(Rha-4),68.4(Rha-5),18.0(Rha-6)。
Forsythoside F(ForsythosideF)
13CNMR(100Mz,DMSO)δ:129.2(C-1),115.5(C-2),144.9(C-3),143.5(C-4),116.4(C-5),119.6(C-6),35.0(C-7),69.4(C-8),125.4(C-1′),114.7(C-2′),145.9(C-3′),148.6(C-4′),115.8(C-5′),121.5(C-6′),145.6(C-7′),113.3(C-8′),166.0(C-9′),102.2(Glc-1),73.0(Glc-2),78.9(Glc-3),70.3(Glc-4),73.2(Glc-5),68.8(Glc-6),101.3(Rha-1),71.7(Rha-2),71.7(Rha-3),76.3(Rha-4),70.4(Rha-5),18.2(Rha-6),103.8(Xyl-1),74.4(Xyl-2)，78.8(Xyl-3)，70.5(Xyl-4)，65.6(Xyl-5)。
Forsythoside I(ForsythosideI)
13CNMR(100Mz,DMSO)δ:129.5(C-1),115.5(C-2),144.7(C-3),143.5(C-4),116.3(C-5),119.5(C-6),35.1(C-7),70.2(C-8),125.6(C-1′),114.9(C-2′),145.3(C-3′),148.2(C-4′),115.7(C-5′),121.3(C-6′),144.9(C-7′),114.7(C-8′),166.1(C-9′),102.7(Glc-1),71.4(Glc-2),77.5(Glc-3),68.1(Glc-4),75.1(Glc-5),66.5(Glc-6),100.7(Rha-1),70.6(Rha-2),70.4(Rha-3),71.9(Rha-4),68.4(Rha-5),17.9(Rha-6)。
Forsythoside J(ForsythosideJ)
13CNMR(100Mz,DMSO)δ:129.2(C-1),115.4(C-2),144.9(C-3),143.5(C-4),116.2(C-5),119.6(C-6),35.0(C-7),69.8(C-8),125.5(C-1′),114.8(C-2′),145.6(C-3′),148.5(C-4′),115.8(C-5′),121.3(C-6′),145.1(C-7′),114.2(C-8′),165.7(C-9′),100.1(Glc-1),73.3(Glc-2),74.1(Glc-3),70.0(Glc-4),75.7(Glc-5),65.7(Glc-6),104.0(Xyl-1),73.3(Xyl-2),76.6(Xyl-3),69.6(Xyl-4),68.2(Xyl-5)。
N-butoxy Forsythoside [(-)-SuspensasideB)
13CNMR(100Mz,DMSO)δ:131.8(C-1),115.0(C-2),146.5(C-3),146.3(C-4),116.5(C-5),119.7(C-6),81.9(C-7),75.4(C-8),127.7(C-1′),115.2(C-2′),145.6(C-3′),148.5(C-4′),115.8(C-5′),121.3(C-6′),145.1(C-7′),114.2(C-8′),167.5(C-9′),100.1(Glc-1),73.3(Glc-2),74.1(Glc-3),70.0(Glc-4),75.7(Glc-5),65.7(Glc-6),104.1(Xyl-1),73.3(Xyl-2),76.6(Xyl-3),69.4(Xyl-4),68.2(Xyl-5)。
Embodiment 1: self-control Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract I GD carbon-13 nmr spectra coupling finger-print
(1) feature extraction thing preparation
Self-control Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract, namely the method preparation of Macroporous Adsorption Resin purifying after adopting alcohol extracting: take Luanchuan green grass or young crops and stick up 20g, add the 20% ethanol refluxing extraction 3 times at 85 DEG C being equivalent to medicinal material 8 volume times amount (160mL), each 1h, filter, merging filtrate, reduced pressure concentration is about extremely without alcohol taste, filter, D101 resin column (its quality is 50g) on filtrate, successively with water, 40% ethanol rushes post (volume of the water and ethanol that rush post is respectively 25 and 40 times of concentrated rear filtrate volume), collect 40% ethanol eluate, evaporated under reduced pressure, obtain homemade Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract 0.78g, it can be used as feature extraction thing.
(2) Luanchuan Qing Qiao lignanoid and the detection of benzyl carbinol glycosides extract I GD carbon-13 nmr spectra finger-print is made by oneself
Get the obtained Luanchuan Qing Qiao lignanoid of step and benzyl carbinol glycosides extract 30mg, be dissolved in 0.5mLDMSO-d 6in, make IGD carbon-13 nmr spectra and detect, obtain Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract I GD carbon-13 nmr spectra finger-print.
(3) Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides feature extraction thing IGD carbon-13 nmr spectra finger-print is made by oneself
1) IGD carbon-13 nmr spectra finger-print is differentiated
In the IGD carbon-13 nmr spectra finger-print of Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract, clearly illustrate the characteristic signal of capsule of weeping forsythia Lignanoids compounds: δ c132.0-136.0,108.0-116.5,144.5-149.0,143.0-146.0,114.0-116.0,118.0-121.0,83.0-88.0,53.0-62.0,68.0-71.0,129.0-133.0,108.0-113.0,146.0-150.0,145.0-154.0,111.0-116.0,117.0-124.0,81.0-82.0 or 84.5-88.0 or 197.0-199.0,49.0-51.0 or 90.0-92.0,68.0-75.0 or 61.0-62.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1,2,3,4,5,6,7,8,9,1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal; In the IGD carbon-13 nmr spectra finger-print of capsule of weeping forsythia lignanoid and benzyl carbinol glycosides extract, clearly illustrate the characteristic signal of capsule of weeping forsythia benzyl carbinol glycosides compounds: δ c129.0-132.0,115.0-117.0,144.0-147.0,143.0-147.0,116.0-118.0,119.0-122.0,34.5-37.0 or 81.0-82.0,69.0-76.0,125.0-128.0,104.0-116.0,145.0-146.0,148.0-150.0,115.0-116.0,121.0-124.0,144.0-148.0,113.0-115.0,165.0-170.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1,2,3,4,5,6,7,8,1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal.Capsule of weeping forsythia lignanoid and benzyl carbinol glycosides compounds: forsythin, table rosin element-4-β-D-Glucose glycosides, loose ester element-β-D-Glucose glycosides, forsythiaside A, Forsythoside H, Forsythoside I etc. all have corresponding NMR signal in IGD carbon-13 nmr spectra finger-print.Accompanying drawing 1-a is shown in by IGD carbon-13 nmr spectra finger-print, and its characteristic peak local widens enlarged drawing and sees accompanying drawing 1-b, 1-c and 1-d.
2) each active component ratio measuring result in Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract is made by oneself as follows:
(4) HPLC method measures forsythin and Forsythoside mass percentage in self-control Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract, and measurement result is as follows:
(5) lignanoid and benzyl carbinol glycosides active component mass percentage measurement result in Luanchuan Qing Qiao lignanoid and benzyl carbinol glycosides extract is made by oneself as follows:
As can be seen from the table, the forsythiaside A content that IGD carbon-13 nmr spectra finger-print calculates is the forsythiaside A content that 18.22%, HPLC measures is 17.44%, and the two error is within 5%.
Embodiment 2: commercially available forsythia suspense extraction IGD carbon-13 nmr spectra coupling finger-print
(1) feature extraction thing is selected
The commercially available forsythia suspense extraction of direct selection (Kang Peiji bio tech ltd, Xi'an) is as feature extraction thing.
(2) commercially available forsythia suspense extraction IGD carbon-13 nmr spectra finger-print detects
Get commercially available forsythia suspense extraction 30mg, be dissolved in 0.5mLDMSO-d 6in, make IGD carbon-13 nmr spectra and detect, obtain commercially available forsythia suspense extraction IGD carbon-13 nmr spectra finger-print.
(3) commercially available forsythia suspense extraction IGD carbon-13 nmr spectra finger-print
1) IGD carbon-13 nmr spectra finger-print is differentiated
In the IGD carbon-13 nmr spectra finger-print of commercially available forsythia suspense extraction, clearly illustrate the characteristic signal of capsule of weeping forsythia Lignanoids compounds: δ c132.0-136.0,108.0-116.5,144.5-149.0,143.0-146.0,114.0-116.0,118.0-121.0,83.0-88.0,53.0-62.0,68.0-71.0,129.0-133.0,108.0-113.0,146.0-150.0,145.0-154.0,111.0-116.0,117.0-124.0,81.0-82.0 or 84.5-88.0 or 197.0-199.0,49.0-51.0 or 90.0-92.0,68.0-75.0 or 61.0-62.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1,2,3,4,5,6,7,8,9,1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal; In the IGD carbon-13 nmr spectra finger-print of commercially available forsythia suspense extraction, clearly illustrate the characteristic signal of capsule of weeping forsythia benzyl carbinol glycosides compounds: δ c129.0-132.0,115.0-117.0,144.0-147.0,143.0-147.0,116.0-118.0,119.0-122.0,34.5-37.0 or 81.0-82.0,69.0-76.0,125.0-128.0,104.0-116.0,145.0-146.0,148.0-150.0,115.0-116.0,121.0-124.0,144.0-148.0,113.0-115.0,165.0-170.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1,2,3,4,5,6,7,8,1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal.Capsule of weeping forsythia lignanoid and benzyl carbinol glycosides compounds: forsythin, loose ester element-β-D-Glucose glycosides, forsythiaside A, Forsythoside H, Forsythoside F, Forsythoside I etc. all have corresponding NMR signal in IGD carbon-13 nmr spectra finger-print.Accompanying drawing 2-a is shown in by IGD carbon-13 nmr spectra finger-print, and its characteristic peak local widens enlarged drawing and sees accompanying drawing 2-b and 2-c.
2) in commercially available forsythia suspense extraction, each active component ratio measuring result is as follows:
(4) HPLC method measures forsythin and Forsythoside mass percentage in commercially available forsythia suspense extraction, and measurement result is as follows:
Forsythin mass percentage W in commercially available forsythia suspense extraction Forsythin（%） 0.67%
Forsythoside mass percentage W in commercially available forsythia suspense extraction Forsythoside（%） 10.34%
(5) in commercially available forsythia suspense extraction lignanoid and benzyl carbinol glycosides active component mass percentage measurement result as follows:
As can be seen from the table, the forsythiaside A content that IGD carbon-13 nmr spectra finger-print calculates is the forsythiaside A content that 10.63%, HPLC measures is 10.34%, and the two error is within 5%.
Embodiment 3: the lignanoid of self-control Luanchuan Folium Forsythia and benzyl carbinol glycosides extract first (precipitation method) IGD carbon-13 nmr spectra coupling finger-print
(1) feature extraction thing preparation
Self-control Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract first, namely the method preparation of poach postprecipitation polishing purification is adopted: take Luanchuan Folium Forsythia 20g, add the water refluxing extraction 2 times at 95 DEG C being equivalent to medicinal material 8 volume times amount (160mL), each 1h, filters, and precipitation is dissolved with MeOH after drying, filter, filtrate dries, and obtains homemade Luanchuan leaf lignanoid and benzyl carbinol glycosides extract 1.56g, it can be used as feature extraction thing.
(2) Folium Forsythia lignanoid and the detection of benzyl carbinol glycosides extract first IGD carbon-13 nmr spectra finger-print is made by oneself
Get the obtained Luanchuan Folium Forsythia lignanoid of step and benzyl carbinol glycosides extract first 30mg, be dissolved in 0.5mLDMSO-d 6in, make IGD carbon-13 nmr spectra and detect, Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract I GD carbon-13 nmr spectra finger-print must be made by oneself.
(3) Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides feature extraction thing first IGD carbon-13 nmr spectra finger-print is made by oneself
1) IGD carbon-13 nmr spectra finger-print is differentiated
In the IGD carbon-13 nmr spectra finger-print of Folium Forsythia lignanoid and benzyl carbinol glycosides extract first, clearly illustrate the characteristic signal of capsule of weeping forsythia Lignanoids compounds: δ c132.0-136.0, 108.0-116.5, 146.0-149.0, 145.0-146.0, 114.0-116.0, 118.0-121.0, 83.0-88.0, 53.0-62.0, 68.0-71.0 or 60.0-62.0, 128.0-133.0, 108.0-113.0, 146.0-150.0, 145.0-154.0, 111.0-116.0, 117.0-124.0, 81.0-82.0 or 84.5-88.0 or 197.0-199.0, 49.0-51.0 or 90.0-92.0, 68.0-75.0 or 61.0-62.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal, in the IGD carbon-13 nmr spectra finger-print of self-control capsule of weeping forsythia lignanoid and benzyl carbinol glycosides extract, clearly illustrate the characteristic signal of capsule of weeping forsythia benzyl carbinol glycosides compounds: δ c129.0-132.0,115.0-117.0,144.0-147.0,143.0-147.0,116.0-118.0,119.0-122.0,34.5-37.0 or 81.0-82.0,69.0-76.0,125.0-128.0,104.0-116.0,145.0-146.0,148.0-150.0,115.0-116.0,121.0-124.0,144.0-148.0,113.0-115.0,165.0-170.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1,2,3,4,5,6,7,8,1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal.Capsule of weeping forsythia lignanoid and benzyl carbinol glycosides compounds: forsythin, Forsythingenin, loose ester element-β-D-Glucose glycosides, "-O-glucoside, forsythiaside A, Forsythoside H, Forsythoside I etc. all have corresponding NMR signal to 1-hydroxyl-rosin element 4 in IGD carbon-13 nmr spectra finger-print.Accompanying drawing 3-a is shown in by IGD carbon-13 nmr spectra finger-print, and its characteristic peak local widens enlarged drawing and sees accompanying drawing 3-b, 3-c and 3-d.
2) each active component ratio measuring result in Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract first is made by oneself as follows:
(4) HPLC method measures forsythin and Forsythoside mass percentage in self-control Folium Forsythia lignanoid and benzyl carbinol glycosides extract first, and measurement result is as follows:
(5) Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract Jia Zhong capsule of weeping forsythia lignanoid and benzyl carbinol glycosides active component mass percentage measurement result is made by oneself as follows:
As can be seen from the table, the forsythiaside A content that IGD carbon-13 nmr spectra finger-print calculates is the forsythiaside A content that 7.87%, HPLC measures is 7.49%, and the two error is within 5%.
Embodiment 4: the lignanoid of self-control Yuzhou Folium Forsythia and benzyl carbinol glycosides extract (resin method) IGD carbon-13 nmr spectra coupling finger-print
(1) feature extraction thing preparation
Self-control Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides extract, namely the method preparation of Macroporous Adsorption Resin purifying after adopting alcohol extracting: take Yuzhou Folium Forsythia 20g, add the water refluxing extraction 3 times at 85 DEG C being equivalent to medicinal material 8 volume times amount (160mL), each 1h, filter, merging filtrate, reduced pressure concentration is about extremely without alcohol taste, filter, D101 resin column (its quality is 50g) on filtrate, successively with water, 40% ethanol rushes post (volume of the water and ethanol that rush post is respectively 25 and 40 times of concentrated rear filtrate volume), collect 40% ethanol eluate, evaporated under reduced pressure, Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides extract 1.79g must be made by oneself, it can be used as feature extraction thing.
(2) Yuzhou Folium Forsythia lignanoid and the detection of benzyl carbinol glycosides extract I GD carbon-13 nmr spectra finger-print is made by oneself
Get the obtained lignanoid of step and benzyl carbinol glycosides extract second 30mg, be dissolved in 0.5mLDMSO-d 6in, make IGD carbon-13 nmr spectra and detect, Folium Forsythia lignanoid and benzyl carbinol glycosides extract second IGD carbon-13 nmr spectra finger-print must be made by oneself.
(3) Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides feature extraction thing IGD carbon-13 nmr spectra finger-print is made by oneself
1) IGD carbon-13 nmr spectra finger-print is differentiated
In the IGD carbon-13 nmr spectra finger-print of self-control Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides extract, clearly illustrate the characteristic signal of capsule of weeping forsythia Lignanoids compounds: δ c132.0-136.0, 108.0-116.5, 144.5-149.0, 143.0-146.0, 114.0-116.0, 118.0-121.0, 83.0-88.0, 53.0-62.0, 68.0-71.0 or 60.0-62.0, 128.0-133.0, 108.0-113.0, 146.0-150.0, 145.0-154.0, 111.0-116.0, 117.0-124.0, 81.0-82.0 or 84.5-88.0 or 197.0-199.0, 49.0-51.0 or 90.0-92.0, 68.0-75.0 or 61.0-62.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal, in the IGD carbon-13 nmr spectra finger-print of self-control Folium Forsythia lignanoid and benzyl carbinol glycosides extract second, clearly illustrate the characteristic signal of capsule of weeping forsythia benzyl carbinol glycosides compounds: δ c129.0-132.0,115.0-117.0,144.0-147.0,143.0-147.0,116.0-118.0,119.0-122.0,34.5-37.0 or 81.0-82.0,69.0-76.0,125.0-128.0,104.0-116.0,145.0-146.0,148.0-150.0,115.0-116.0,121.0-124.0,144.0-148.0,113.0-115.0,165.0-170.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1,2,3,4,5,6,7,8,1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal.Capsule of weeping forsythia lignanoid and benzyl carbinol glycosides compounds: forsythin, loose ester element-β-D-Glucose glycosides, "-O-glucoside, forsythiaside A, Forsythoside H, Forsythoside I etc. all have corresponding NMR signal to 1-hydroxyl-rosin element 4 in IGD carbon-13 nmr spectra finger-print.Accompanying drawing 4-a is shown in by IGD carbon-13 nmr spectra finger-print, and its characteristic peak local widens enlarged drawing and sees accompanying drawing 4-b, 4-c and 4-d.
2) each active component ratio measuring result in Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides extract is made by oneself as follows:
(4) HPLC method measures forsythin and Forsythoside mass percentage in self-control Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides extract, and measurement result is as follows:
(5) capsule of weeping forsythia lignanoid and benzyl carbinol glycosides active component mass percentage measurement result in Yuzhou Folium Forsythia lignanoid and benzyl carbinol glycosides extract is made by oneself as follows:
As can be seen from the table, the Determination of forsythin that IGD carbon-13 nmr spectra finger-print calculates is the forsythiaside A content that 6.99%, HPLC measures is 6.70%, and the two error is within 5%.
Embodiment 5: self-control Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second (resin method) IGD carbon-13 nmr spectra coupling finger-print
(1) feature extraction thing preparation
Self-control Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second, namely the method preparation of Macroporous Adsorption Resin purifying after adopting alcohol extracting: take Luanchuan Folium Forsythia 20g, add the water refluxing extraction 3 times at 85 DEG C being equivalent to medicinal material 8 volume times amount (160mL), each 1h, filter, merging filtrate, reduced pressure concentration is about extremely without alcohol taste, filter, D101 resin column (its quality is 50g) on filtrate, successively with water, 40% ethanol rushes post (volume of the water and ethanol that rush post is respectively 30 and 40 times of concentrated rear filtrate volume), collect 40% ethanol eluate, evaporated under reduced pressure, Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second 1.56g must be made by oneself, it can be used as feature extraction thing.
(2) Luanchuan Folium Forsythia lignanoid and the detection of benzyl carbinol glycosides extract second IGD carbon-13 nmr spectra finger-print is made by oneself
Get the obtained lignanoid of step and benzyl carbinol glycosides extract second 30mg, be dissolved in 0.5mLDMSO-d 6in, make IGD carbon-13 nmr spectra and detect, Folium Forsythia lignanoid and benzyl carbinol glycosides extract second IGD carbon-13 nmr spectra finger-print must be made by oneself.
(3) Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides feature extraction thing second IGD carbon-13 nmr spectra finger-print is made by oneself
1) IGD carbon-13 nmr spectra finger-print is differentiated
In the IGD carbon-13 nmr spectra finger-print of self-control Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second, clearly illustrate the characteristic signal of capsule of weeping forsythia Lignanoids compounds: δ c132.0-136.0, 108.0-116.5, 144.5-149.0, 143.0-146.0, 114.0-116.0, 118.0-121.0, 83.0-88.0, 53.0-62.0, 68.0-71.0 or 60.0-62.0, 128.0-133.0, 108.0-113.0, 146.0-150.0, 145.0-154.0, 111.0-116.0, 117.0-124.0, 81.0-82.0 or 84.5-88.0 or 197.0-199.0, 49.0-51.0 or 90.0-92.0, 68.0-75.0 or 61.0-62.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal, in the IGD carbon-13 nmr spectra finger-print of self-control Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second, clearly illustrate the characteristic signal of capsule of weeping forsythia benzyl carbinol glycosides compounds: δ c129.0-132.0,115.0-117.0,144.0-147.0,143.0-147.0,116.0-118.0,119.0-122.0,34.5-37.0 or 81.0-82.0,69.0-76.0,125.0-128.0,104.0-116.0,145.0-146.0,148.0-150.0,115.0-116.0,121.0-124.0,144.0-148.0,113.0-115.0,165.0-170.0 to be respectively on lignanoid and benzyl carbinol glycosides skeleton 1,2,3,4,5,6,7,8,1 ', 2 ', 3 ', 4 ', 5 ', 6 ', 7 ', 8 ', 9 ' position carbon signal.Capsule of weeping forsythia lignanoid and benzyl carbinol glycosides compounds: forsythin, loose ester element-β-D-Glucose glycosides, forsythiaside A, Forsythoside H, Forsythoside I etc. all have corresponding NMR signal in IGD carbon-13 nmr spectra finger-print.Accompanying drawing 5-a is shown in by IGD carbon-13 nmr spectra finger-print, and its characteristic peak local widens enlarged drawing and sees accompanying drawing 5-b, 5-c and 5-d.
2) each active component ratio measuring result in Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second is made by oneself as follows:
(4) HPLC method measures forsythin and Forsythoside mass percentage in self-control Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract second, and measurement result is as follows:
(5) Luanchuan Folium Forsythia lignanoid and benzyl carbinol glycosides extract Yi Zhong capsule of weeping forsythia lignanoid and benzyl carbinol glycosides active component mass percentage measurement result is made by oneself as follows:
As can be seen from the table, the Determination of forsythin that IGD carbon-13 nmr spectra finger-print calculates is the forsythiaside A content that 7.84%, HPLC measures is 7.69%, and the two error is within 5%.

Claims (8)

1. differentiate a method for capsule of weeping forsythia spin-off, comprise the following steps:
1) capsule of weeping forsythia spin-off is got directly as the capsule of weeping forsythia lignanoid containing active component group and benzyl carbinol glycosides feature extraction thing;
2) carry out IGD carbon-13 nmr spectra finger-print to described capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing to detect, obtain several active component characteristic peak peak intensities in feature extraction thing according to finger-print; And determine the characteristic peak peak intensity of described each active component respective standard with reference to product by same way;
In capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing, the characteristic peak of lignanoids active component is C-7 absorption peak, and its chemical shift is δ c83.0 ~ 88.0; Select C-7 ' distinguish the forsythin in lignanoids active component further as characteristic peak and show rosin element-4-β-D-Glucose glycosides, their chemical shift is δ c81.0 ~ 82.0;
In capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing, the characteristic peak of benzyl carbinol glycosides active component is C-9 ' absorption peak, and its chemical shift is δ c165.0 ~ 170.0; Select C-1 or C-6 to distinguish forsythiaside A in benzyl carbinol glycosides active component and Forsythoside H further as characteristic peak, their chemical shift is respectively δ c119.0 ~ 120.0 and δ c129.0 ~ 130.0;
The standard of the active component in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing is forsythin or forsythiaside A with reference to product;
3) absolute content of described standard with reference to product is obtained by high-efficient liquid phase technique;
When differentiating capsule of weeping forsythia lignanoids active component, it is acetonitrile that the condition of described high-efficient liquid phase technique comprises mobile phase: water=(20 ~ 30): (70 ~ 80);
When differentiating benzyl carbinol glycosides active component, it is acetonitrile that the condition of described high-efficient liquid phase technique comprises mobile phase: glacial acetic acid solution=(10 ~ 20): (80 ~ 90);
4) utilize the ratio of the characteristic peak peak intensity of each active component characteristic peak peak intensity and respective standard reference product and the absolute content of described standard reference product, calculate the content of each active component in capsule of weeping forsythia spin-off and the content of active component group.
2. method according to claim 1, is characterized in that, adopts and has the extraction process that the obtains clear IGD carbon-13 nmr spectra finger-print extracting mode as described capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing.
3. method according to claim 1, it is characterized in that, step 2) in, carry out IGD carbon-13 nmr spectra finger-print to capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing to detect, the solvent dissolving capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing is deuterated dimethyl sulfoxide or deuterated methanol.
4., according to the arbitrary described method of claims 1 to 3, it is characterized in that, step 2) in, according to size and the position of characteristic peak peak intensity, several active components in capsule of weeping forsythia lignanoid and benzyl carbinol glycosides feature extraction thing are sorted.
5. method according to claim 4, is characterized in that, when differentiating capsule of weeping forsythia lignanoids active component, it is acetonitrile that the condition of described high-efficient liquid phase technique comprises mobile phase: water=25:75; Determined wavelength is 277nm;
When differentiating benzyl carbinol glycosides active component, it is acetonitrile that the condition of described high-efficient liquid phase technique comprises mobile phase: glacial acetic acid solution=15:85; Determined wavelength is 330nm.
6. the method according to claim 1 or 2 or 3 or 5, is characterized in that, step 3) in, with reference to the absolute content of product, described standard refers to that the standard using quantitative test means mensuration is with reference to the mass percentage of product.
7. want the method described in 6 according to right, it is characterized in that, step 4) in, the coupling formula calculating the content of each active component is:
W n = W 1 M n h n M 1 h 1 ; Wherein:
W 1for step 3) a certain active component is corresponding in the capsule of weeping forsythia spin-off that measures by quantitative test means standard is with reference to the absolute content of product;
M 1for standard corresponding to a certain active component in described capsule of weeping forsythia spin-off is with reference to the carbon number that molecular weight/quantitatively peak is corresponding of product;
H 1for standard corresponding to a certain active component in the capsule of weeping forsythia spin-off by IGD carbon-13 nmr spectra determining fingerprint pattern is with reference to the characteristic peak peak intensity of product;
W nfor the mass percentage of active component a certain in capsule of weeping forsythia spin-off;
M nfor the carbon number that molecular weight/quantitatively peak is corresponding of active component a certain in capsule of weeping forsythia spin-off;
H nfor the characteristic peak peak intensity of a certain active component in the capsule of weeping forsythia spin-off by IGD carbon-13 nmr spectra determining fingerprint pattern.
8. the method according to claim 1 or 2 or 3 or 5 or 7, it is characterized in that, described capsule of weeping forsythia spin-off comprises: forsythia suspense extraction or capsule of weeping forsythia natural drug.
CN201410044108.0A 2013-01-30 2014-01-30 A kind of method differentiating capsule of weeping forsythia spin-off Active CN103808749B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201410044108.0A CN103808749B (en) 2013-01-30 2014-01-30 A kind of method differentiating capsule of weeping forsythia spin-off

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN201310036486 2013-01-30
CN201310036486X 2013-01-30
CN201310036486.X 2013-01-30
CN201410044108.0A CN103808749B (en) 2013-01-30 2014-01-30 A kind of method differentiating capsule of weeping forsythia spin-off

Publications (2)

Publication Number Publication Date
CN103808749A CN103808749A (en) 2014-05-21
CN103808749B true CN103808749B (en) 2016-04-13

Family

ID=50705832

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410044108.0A Active CN103808749B (en) 2013-01-30 2014-01-30 A kind of method differentiating capsule of weeping forsythia spin-off

Country Status (1)

Country Link
CN (1) CN103808749B (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105287610B (en) * 2014-06-27 2021-11-16 石家庄以岭药业股份有限公司 Application of forsythoside I and preparation method thereof
CN104698022B (en) * 2015-04-02 2016-09-28 河南中医学院 A kind of method measuring arylnaphthalene lactone type lignanoid purity based on quantitative hydrogen nuclear magnetic resonance spectroscopy
CN112946113B (en) * 2021-02-01 2022-09-30 湖北医药学院 Method for identifying genuine medicinal material Shiweir fructus forsythiae basal source
CN114166980A (en) * 2021-12-28 2022-03-11 山西振东泰盛制药有限公司 Method for constructing lignan fingerprint in ginkgo leaf medicinal material, extract and single preparation thereof

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2324182C1 (en) * 2006-11-13 2008-05-10 Государственное образовательное учреждение высшего профессионального образования "Санкт-Петербургский государственный медицинский университет имени академика И.П. Павлова Федерального агентства по здравоохранению и социальному развитию" Method of diagnostics of microfloral abnormality of oral cavity
CN101152261A (en) * 2007-09-04 2008-04-02 北京理工大学 Application of phenyl ethyl alcohol glycosides and xylogen glycosides component in forsythia (fruit or leaf)
CN101613384B (en) * 2008-06-25 2011-12-07 中国科学院大连化学物理研究所 Dihydroxyphenethyl alcohol glycoside compound and preparation method thereof
CN102749348B (en) * 2012-06-18 2014-06-11 河南省科高植物天然产物开发工程技术有限公司 Method for identifying active components in medicinal plant

Also Published As

Publication number Publication date
CN103808749A (en) 2014-05-21

Similar Documents

Publication Publication Date Title
CN105606734B (en) A kind of quick high separation liquid chromatographic detection honeysuckle and the method for Honeysuckle flower medicinal material
CN102749348B (en) Method for identifying active components in medicinal plant
CN103808749B (en) A kind of method differentiating capsule of weeping forsythia spin-off
CN105891353A (en) Phlegm heat removing capsule fingerprint spectrum and building method thereof
CN103674996B (en) A kind of method differentiating red rooted salvia or spin-off
CN103808751B (en) A kind of method differentiating traditional Chinese medicine honeysuckle or spin-off
CN104374854B (en) A kind of method of multiple phenolic content in HPLC wavelength handoff technique Simultaneously test Noni juice
Pfoze et al. Isolation of protoberberine alkaloids from stem bark of Mahonia manipurensis Takeda using RP-HPLC
Li et al. Quality evaluation of Radix Stemonae through simultaneous quantification of bioactive alkaloids by high‐performance liquid chromatography coupled with diode array and evaporative light scattering detectors
CN107561192A (en) R accuses the method accused according to spring and S according to spring content in a kind of measure chromatogram of Radix Isatidis and its product
CN101791366A (en) Method for testing quality of Discorea nipponica Makino in different places and medicinal materials of same genera
CN109991327A (en) One surveys the methods for commenting method evaluation field thistle quality more
CN103868943B (en) A kind of method of differentiating Fructus Ligustri Lucidi or spin-off
CN103134877A (en) Method measuring C6-C3 type phenolic acids compound content in tobacco
CN103134827B (en) Method for identifying angled bittersweet seed medicinal materials
CN112684036A (en) Fingerprint spectrum determination method of kidney-tonifying capsules containing leeches and application of fingerprint spectrum determination method
CN104215614B (en) A kind of high performance liquid chromatography-diode array/fluorescence detector Series detectors barbaloin A, B and the method for aloe-emodin
CN103048347B (en) Method for distinguishing root-bark medicinal materials of celastrus angulatus
CN103105408B (en) A kind of method differentiating Fructus Corni or spin-off
CN109991328A (en) A kind of affine cudweed one surveys the quality evaluating method more commented
CN107449848A (en) A kind of method of La Ruia extracts in discriminating cosmetic material
Wang et al. HPLC fingerprint combined with quantitation of main effective components and chemometrics as an efficient method for quality evaluation of Oviductus Ranae
CN107764924A (en) The detection method of active ingredient in asthma particle
CN103808750B (en) A kind of method of differentiating black false hellebore medicinal material
CN106226409A (en) Clear cloperastine breathes heavily the detection method of granule

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20230620

Address after: No. 6 Shangding Road, Jinshui District, Zhengzhou City, Henan Province, 450000

Patentee after: Wang Shuncai

Address before: 450002 Henan city of Zhengzhou Province Red Road No. 58

Patentee before: HENAN BRANCH OF PLANT NATURAL PRODUCTS OF HIGH ENGINEERING TECHNOLOGY DEVELOPMENT CO.,LTD.