CN102907425A

CN102907425A - Celangulin mother liquor and preparation method and quality inspection method thereof

Info

Publication number: CN102907425A
Application number: CN2012103747900A
Authority: CN
Inventors: 赵天增; 张海艳; 董建军; 范毅; 于立芹; 魏悦; 刘雨晴; 郭唯; 常霞; 陈玲; 李倩
Original assignee: BIOLOGICAL DEVELOPING CENTER OF HENAN ACADEMY OF SCIENCES; Henan Kegao Vegetable Natural Product Development Engineering Technology Co ltd
Current assignee: BIOLOGICAL DEVELOPING CENTER OF HENAN ACADEMY OF SCIENCES; Henan Kegao Vegetable Natural Product Development Engineering Technology Co ltd
Priority date: 2012-09-29
Filing date: 2012-09-29
Publication date: 2013-02-06
Anticipated expiration: 2032-09-29
Also published as: CN102907425B

Abstract

The invention relates to celangulin mother liquor and emulsion in water, a preparation method and a quality inspection method thereof. The celangulin mother liquor comprises, by mass, active ingredients of 1.9-3.5% of angulatin A, 3.0-5.0% of angulatin B, 2.5-4.0% of angulatin G and 2.50-4.10% of angulatin E. The celangulin mother liquor is prepared by an ultrasonic recycling extraction method, and the quality inspection method adopts the IGD carbon-13 nuclear magnetic resonance spectroscopy coupling fingerprint technology. The celangulin mother liquor content is increased greatly, and accuracy, stability, repeatability and feasibility of the quality inspection method are high.

Description

A kind of Celastrus angulatus mother liquor and preparation method thereof and quality determining method

Technical field

The invention belongs to the botanical pesticide technical field, particularly, related to a kind of Celastrus angulatus mother liquor and aqueous emulsion thereof, and their preparation method and quality determining method.

Background technology

Botanical pesticide belongs to a branch in the biopesticide category, is to implement the effects such as desinsection, sterilization as active component with certain contained in plant class stable elements.Because these active components derive from plant itself, are easy to natural decomposition; And the diversity with structure, pest are difficult to develop immunity to drugs.Therefore, botanical pesticide is one of main direction of from now on countries in the world Development in Green Pesticides.Domestic botanical pesticide comprise Dalmatian chrysanthemum, margosine, matrine, nicotinic insecticide and Celastrus angulatus agrochemical [1. Wei Yan is quick. biopesticide and application technology question and answer thereof. China Agricultyre University Press 2007; 2. Hou Taiping. Research Progresses on Botanical Pesticides. Chinese agriculture science and technology Leader 2006,8 (6): 12].

Celastrus angulatus (Celastrus Angulatus Marim) is the perennial fallen leaves rattan of Celastraceae (Celastraceae) Celastrus (Celastrus) shape shrub plant, have another name called bitter bark, red, the wintercreeper euonymus of frost, cabbageworm medicine, radish medicine fall, its root skin powder, leaf powder and stem skin powder have long prevention and elimination of disease and pests historical China is among the people, that China has one of minority pesticide plant of potentiality to be exploited [Lu Lingxian most, Deng. Insecticidal Plant Celastrus Angulatus is introduced a fine variety the breeding First Report of Studies. the Hubei forestry science and technology, 1987,3:8].Celastrus angulatus has ample natural resources in command in China, in Henan, the territories, mountain, more than 10 province hills such as Shaanxi, Hubei all have the growth of distribution [Wu Wenjun. the Insecticidal Plant Celastrus Angulatus research overview. plant protection, 1993,3 (17): 34.].The insecticidal activity research of relevant Celastrus angulatus has more report, and confirmed that the Celastrus angulatus extract has that insecticidal activity is strong, the characteristics [Ke Zhiguo such as be difficult for developing immunity to drugs, Deng. pesticide plant---the research of Celastrus angulatus seed. the 3rd the academic Conference Papers collection of Hubei And Hunan plant protection agricultural chemicals, 2004,294.].From Celastrus angulatus, isolated plurality of active ingredients, and clear and definite its has various biologically actives such as food refusal, anesthesia, poisoning to insect, can be used to prevent and treat cabbage caterpillar on the vegetables, rice plant skipper, paddy rice mythimna separata, blister beetle sawfly larva, pest of stored grain corn weevil etc. [Wu Wenjun. the research of vegetable insecticide Celastrus angulatus with use. Chemical Industry Press, 2010.].At present, the main liquid dosage form of Celastrus angulatus insecticide has 1% Celastrus angulatus missible oil [CN94103655.3; The formal agriculture chemical registration of 1% Celastrus angulatus missible oil card (PD20101574)], 0.2% Celastrus angulatus missible oil [CN92113104.6], 0.5% Celangulins ME [CN99109275.9], 6% Celastrus angulatus mother liquor [the formal agriculture chemical registration card of 6% Celastrus angulatus mother liquor (PD20101575)] etc.

The extraction separation method of the Celastrus angulatus that bibliographical information and existing product adopt is the conventional methods such as solvent reflux extraction, percolation, and these methods are consuming time bothersome, and recovery rate is low.Ultrasonic technique is as a kind of effective extracting method, be widely used in [Fu Guixiang in the extraction research of natural products, Deng. assay and the comparison of not of the same race and different harvest time Total Flavone in Hippopae. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 1997,22 (3): 147～148.].Ultrasonic circulation technique is to apply hyperacoustic natural product extraction new technology [Zheng Mingyu in the process that feed liquid circulates; Zheng Lingling; Wen Liying; Wu Yingzi; Guo Jianpeng; Geranium wilfordii tannin ultrasonic circulation Study on extraction Chinese experimental pharmacology of traditional Chinese medical formulae magazine, 2010,16 (12): 7.], have and extract the unique advantage that temperature is low, extraction efficiency is high, extraction time is short.We have carried out deep, systematic research [Hai-Lin Qin, et al.ANew Sesquiterpene from Celastrus Angulatus.Chin Chem Lett, 1999,10 (10): 825-826 to wild Celastrus angulatus; Hai-Yan Zhang, et al.Two new sesquiterpene polyol esters from the root barks of Celastrus angulatus.Journal of asian natural products research, 2011,13 (4): 304.], find a series of active insecticidal components and determined its chemical constitution, and through ultrasonic circulation extraction insecticide active substance wherein, make environment friendly agricultural Celastrus angulatus mother liquor.

The quality of Celastrus angulatus does not lie in certain single-activity composition, and its effect is the result of the collaborative proportioning of a plurality of active components.Studies have shown that in the Celastrus angulatus that main active insecticidal components is polyol ester compound and the alkaloid thereof with β-dihydroagarofuran sequiterpene skeleton, the general designation Celastrus angulatus.Celastrus angulatus contains take Celangulin A, Celangulin B as main multiple insecticidal constituent.At present, the detection method of Celastrus angulatus component in the Celastrus angulatus mother liquor: the one, measure the benzoic acid that produces after the hydrolysis of Celastrus angulatus mother liquor, operating procedure is extremely complicated, and the result is unreliable, and feasibility is poor; The 2nd, only one of them component Celangulin A (or Celangulin V) is carried out quantitative analysis [Wu Wenjun, Deng. the research and development .Pesticides (agricultural chemicals) of vegetable insecticide 0.2% Celastrus angulatus missible oil, 2001,40 (3): 17.], this quality testing pattern of carrying out quantitative analysis for single component can not effectively be controlled the inherent quality of Celastrus angulatus mother liquor, can't satisfy current for the objective an urgent demand of effectively estimating and controlling the Celastrus angulatus product quality.Fingerprint pattern technology oneself become internationally recognized difference estimate the most effective means of natural plant product and raw material thereof [Luo Guoan, etc. the classification of traditional Chinese medicine fingerprint and development. Chinese Journal of New Drugs, 2002,11 (1): 46.].Have no report about Celastrus angulatus mother liquor and aqueous emulsion finger-print thereof at present.

IGD carbon-13 nmr spectra coupling (IGD ¹³C NMR coupling) fingerprint pattern technology also is inverted gated decoupling carbon-13 nmr spectra coupling fingerprint pattern technology, be study for many years proton nmr spectra ( ¹H NMR) fingerprint pattern technology [Zhao Tianzeng, etc. ¹HNMR fingerprint technique plant identification Chinese medicine. Chinese herbal medicine, 2000,31 (11): 868-870.] unite a kind of new comprehensive fingerprint pattern technology of non-single means that other technologies (for example present most widely used high efficiency liquid phase (HPLC) fingerprint pattern technology) propose on the basis.

Day by day be subject to the great attention of country and society along with medicine, food security, the Celastrus angulatus that can be widely used in the control of insect of pollution-free vegetable, crops and non-crop class is in great demand, for the application of IGD carbon-13 nmr spectra coupling fingerprint pattern technology provides wide basis.Research and the application of Celastrus angulatus mother liquor and aqueous emulsion IGD carbon-13 nmr spectra coupling fingerprint pattern technology thereof, not only can solve the wherein qualitative question of active component, also for strengthening systematization and the standardization of its inherent composition Study, accelerate the modern development of botanical pesticide Celastrus angulatus, realize the assurance that science is provided in line with international standards.Along with this technology applying in other traditional Chinese medicines and extract thereof, botanical pesticide, the great scientific value of this technology will be increasingly outstanding.

Summary of the invention

The problem that exists in order to solve the aspects such as botanical pesticide prior art, quality the object of the invention is to, and a kind of good botanical pesticide Celastrus angulatus mother liquor and aqueous emulsion thereof are provided.

Another object of the present invention is to, the preparation method of a kind of described Celastrus angulatus mother liquor and aqueous emulsion thereof is provided.

Another purpose of the present invention is, the quality determining method of a kind of described Celastrus angulatus mother liquor and aqueous emulsion thereof is provided.

For achieving the above object, a kind of Celastrus angulatus mother liquor provided by the invention wherein contains the active component of following quality percentage composition: Celangulin A1.9 ~ 3.5%, Celangulin B3.0 ~ 5.0%, Celangulin G2.5 ~ 4.0%, Celangulin E2.50 ~ 4.10%.

Celastrus angulatus mother liquor provided by the invention, preferably, the active component that wherein also contains following quality percentage composition: Celangulin C, Celastrus angulatus II and Celangulin F1.2 ~ 3.5%, Celastrus angulatus IV0.3 ~ 1.0%, Celangulin A, Celangulin E and Celastrus angulatus C(celangulatinC) 3.0 ~ 4.5%, Celangulin H0.3 ~ 2.0%.

Described active component in the Celastrus angulatus mother liquor of the present invention is 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound.

Namely, the present invention's Celastrus angulatus mother liquor provided by the invention, the active component that wherein contains following quality percentage composition: Celangulin A1.9 ~ 3.5%, Celangulin B3.0 ~ 5.0%, Celangulin C, Celastrus angulatus II and Celangulin F1.2 ~ 3.5%, Celangulin G2.5 ~ 4.0%, Celastrus angulatus IV0.3 ~ 1.0%, Celangulin A, Celangulin E and Celastrus angulatus C(celangulatinC) 3.0 ~ 4.5%, Celangulin H0.3 ~ 2.0%.

Wherein, the quality percentage composition of Celangulin C is 0.35 ~ 1.40%, the quality percentage composition of Celangulin F is 0.10 ~ 0.90%, and the quality percentage composition of Celastrus angulatus II is 0.60 ~ 1.21%, Celastrus angulatus C(celangulatin C) the quality percentage composition be 0.30 ~ 0.55%.

The total quality percentage composition of each active component in the Celastrus angulatus mother liquor of the present invention (4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound) is not less than 10%.

Further, the total quality percentage composition of each active component is 11.5 ~ 18.5% in the Celastrus angulatus mother liquor.

Further, kind is that the total quality percentage composition of active component is 16.3 ~ 18.4% in the Celastrus angulatus mother liquor of Xichuan County, henan Province, and kind is that the total quality percentage composition of active component is 11.7 ~ 13.2% in the Celastrus angulatus mother liquor of Baoji.

The preparation method of Celastrus angulatus mother liquor provided by the invention comprises: taking by weighing the Celastrus angulatus root or/and skin is pulverized, is that 5 ~ 7 times benzene extracts 1 ~ 2 time in 45 ~ 55 ℃ of lower ultrasonic circulations with volume, and reduced pressure concentration became medicinal extract after filtrate merged, and namely got the Celastrus angulatus mother liquor.

Further, the Celastrus angulatus root is or/and skin is crossed 10 ~ 24 mesh sieves, preferred 24 orders after pulverizing.

Further, when ultrasonic circulation extracts, extracted preferred 55 minutes 35 ~ 55 minutes at every turn.

Further, the temperature extracted of ultrasonic circulation is preferred 50 ℃.

Further, the density of concentrated medicinal extract is 1.1 ~ 1.3.

The Celastrus angulatus root is or/and the active component in the skin except the described composition of Celastrus angulatus mother liquor of the present invention, also comprises Celangulin U, X; Celangulin P, Celangulin V II, Celastrus angulatus XIX; The 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compounds such as Celangulin J, Celastrus angulatus III, and the composition of other classifications such as flavones, alkaloid, triterpene.

A kind of Celastrus angulatus aqueous emulsion provided by the invention comprises described Celastrus angulatus mother liquor.

Described Celastrus angulatus aqueous emulsion provided by the invention comprises the composition of following weight portion: 15 ~ 20 parts in described Celastrus angulatus mother liquor, 8 ~ 12 parts of emulsifier, 3 ~ 4 parts of antifreezes.

Wherein, described emulsifier is: alkylaryl polyoxyethylene poly-oxygen propylene aether or alkylaryl polyoxyethylene APEO, preferred alkyl aryl polyoxyethylene poly-oxygen propylene aether.

Wherein, described antifreeze is: ethylene glycol or propane diols, preferred ethylene glycol.

A kind of Celastrus angulatus aqueous emulsion provided by the invention also comprises solvent.

Wherein, solvent is water.

Celastrus angulatus aqueous emulsion of the present invention, auxiliary material wherein, namely emulsifier, antifreeze and solvent also can be conventional kind and the consumption of this area.Can also make used additives, also be that the routine of this area is selected.

Total quality percentage composition of each active component in the Celastrus angulatus aqueous emulsion of the present invention (4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound) is not less than 2%.

Further, the total quality percentage composition of each active component is 2.3 ~ 3.0% in the Celastrus angulatus aqueous emulsion.

Further, kind is that the total quality percentage composition of active component is 2.7 ~ 3.0% in the Celastrus angulatus mother liquor of Xichuan County, henan Province, and kind is that the total quality percentage composition of active component is 2.3 ~ 2.7% in the Celastrus angulatus mother liquor of Baoji.

The preparation method of described Celastrus angulatus aqueous emulsion provided by the invention comprises: described Celastrus angulatus mother liquor is mixed in described ratio with emulsifier, antifreeze and solvent, and get final product.

The quality determining method of Celastrus angulatus mother liquor provided by the invention may further comprise the steps:

1) the Celastrus angulatus mother liquor is extracted, obtain containing the Celastrus angulatus mother liquor feature extraction thing of active component group;

2) Celastrus angulatus mother liquor feature extraction thing is carried out IGD carbon-13 nmr spectra finger-print and detect, obtain several active component characteristic peak peak intensities in the described Celastrus angulatus mother liquor feature extraction thing according to finger-print; And determine the characteristic peak peak intensity of the corresponding criterion referenced product of described each active component with same way as (IGD carbon-13 nmr spectra finger-print);

3) measure the absolute content of the criterion referenced product described in the Celastrus angulatus mother liquor that obtain by the quantitative analysis means;

4) utilize ratio and the described absolute content of described characteristic peak peak intensity (the characteristic peak peak intensities of each active component characteristic peak peak intensity and corresponding criterion referenced product), calculate the content of each active component in the Celastrus angulatus mother liquor and the total content of this active component, i.e. the content of active component group.

Wherein, in the step 1), the preparation method of Celastrus angulatus mother liquor feature extraction thing comprises: take by weighing the Celastrus angulatus mother liquor, filter rear at 55 ~ 65 ℃ of lower reduced pressure concentrations, reclaim solvent to the medicinal extract shape, medicinal extract with 6 ~ 10 times of amount volumes, the dissolving of 80% ~ 90% methyl alcohol after, use again the petroleum ether extraction 1 ~ 2 time of 3 ~ 5 times of amount volumes, reduced pressure concentration after methanol layer filters, reclaim solvent to doing, namely get Celastrus angulatus mother liquor feature extraction thing.

Further, use petroleum ether extraction, extract 20 ~ 40 minutes at every turn, preferred 30 minutes.

Further, after the Celastrus angulatus mother liquor filters preferably at 60 ℃ of lower reduced pressure concentrations.

The processing of Celastrus angulatus mother liquor feature extraction thing and detection method are as follows: get Celastrus angulatus mother liquor feature extraction thing, be dissolved in CDCl ₃In, make IGD carbon-13 nmr spectra finger-print and detect.

Wherein, step 2) in, the active component characteristic peak in the described Celastrus angulatus mother liquor feature extraction thing is: C-15 absworption peak, its chemical shift are δ _C60.0 ~ 66.0.

Further, select C-8 further to distinguish Celangulin A, Celangulin E, Celastrus angulatus C as characteristic peak, its (C-8) chemical shift is δ _C73.3 ~ 74.7; Select C-11 further to distinguish Celangulin C, Celangulin F and Celastrus angulatus II as characteristic peak, its (C-11) chemical shift is δ _C82.5 ~ 83.9.

Wherein, step 2) in, described peak intensity can adopt peak height method, area integral method or gravimetric method to calculate.

Wherein, in the step 3), the absolute content of described criterion referenced product refers to: use the Celastrus angulatus mother liquor Plays of quantitative analysis means mensuration with reference to the quality percentage composition of product.

Wherein, described quantitative analysis means are high performance liquid chromatography (HPLC) method.

Further, the condition of HPLC method is: chromatographic column is take octadecyl silane as filler, and mobile phase is acetonitrile: the mixed solvent of water=(60:40) ~ (55:45), and flow velocity is 1mL/min, the detection wavelength is 232nm.Acetonitrile: the preferred 55:45 of water.

Wherein, described criterion referenced product are Celangulin A.

Wherein, the calculating of the total content of the content of each active component and this active component is with IGD carbon-13 nmr spectra and the coupling of quantitative analysis means by the coupling computing formula in the Celastrus angulatus mother liquor, be in the step 4), the coupling formula that calculates the content of each active component is:

X_{n} = \frac{X_{1} M_{n} h_{n}}{M_{1} h_{1}};

Wherein:

X ₁Be the absolute content (quality percentage composition) of step 3) with the criterion referenced product that a certain active component is corresponding in the Celastrus angulatus mother liquor of quantitative analysis means mensuration;

M ₁Molecular weight for criterion referenced product corresponding to described a certain active component;

h ₁Be the characteristic peak peak intensity by criterion referenced product corresponding to a certain active component described in the Celastrus angulatus mother liquor feature extraction thing of IGD carbon-13 nmr spectra determining fingerprint pattern;

X _nQuality percentage composition for a certain active component in the Celastrus angulatus mother liquor;

M _nMolecular weight for a certain active component;

h _nBe the characteristic peak peak intensity by a certain active component in the Celastrus angulatus mother liquor feature extraction thing of IGD carbon-13 nmr spectra determining fingerprint pattern.

Also can calculate coefficient and the overall coefficient of each active component in the Celastrus angulatus mother liquor by the coupling formula.

Further, the computing formula of described coefficient is:

X_{n} = \frac{X_{1} M_{n} h_{n}}{M_{1} h_{1}};

F wherein _nRatio coefficient for the criterion referenced product quality percentage composition that a certain active component is corresponding with it in the Celastrus angulatus mother liquor.M ₁, h ₁, M _nAnd h _nImplication with calculating the corresponding definition in each active component content coupling formula in the Celastrus angulatus mother liquor.Overall coefficient refers to the coefficient sum of each composition.

This coefficient F _nBe applicable to too calculate active component and active component group in the Celastrus angulatus aqueous emulsion.

The quality determining method of Celastrus angulatus aqueous emulsion provided by the invention utilizes said method to detect the content that obtains certain each active component in the Celastrus angulatus mother liquor, extrapolates the quality percentage composition of each active component in the Celastrus angulatus aqueous emulsion again.The Celastrus angulatus aqueous emulsion here is to be prepared by the Celastrus angulatus mother liquor.

Wherein, the cubage formula of each active component is in the Celastrus angulatus aqueous emulsion:

W _n=W ₁F _nWherein:

W _nBe a certain active component quality percentage composition in the Celastrus angulatus aqueous emulsion;

W ₁Absolute content (quality percentage composition) for the criterion referenced product that a certain active component is corresponding in the Celastrus angulatus aqueous emulsion of measuring with the quantitative analysis means.

Wherein, described quantitative analysis means are high performance liquid chromatography.

The total content of this active component is exactly the W of similar each active component in Celastrus angulatus mother liquor or the Celastrus angulatus aqueous emulsion _nSum, i.e. the content of active component group, single active series of compounds is counted F _nSum is the Celastrus angulatus overall coefficient.

Quality determining method of the present invention can be used for detecting the present invention 10% Celastrus angulatus mother liquor, 6% Celastrus angulatus mother liquor or the Celastrus angulatus mother liquor of other concentration, and the present invention 2% Celastrus angulatus aqueous emulsion, 1% Celastrus angulatus aqueous emulsion or the Celastrus angulatus aqueous emulsion of other concentration.

The described active component group of the inventive method is especially carried out the Celastrus angulatus medicinal material after ultrasonic circulation extracts the Celastrus angulatus mother liquor that obtains and the active component group in the Celastrus angulatus aqueous emulsion.

Celastrus angulatus medicinal material of the present invention refers to that the root of Celastrus angulatus plant is or/and the position of skin.

The calculating of the content of each active component of the present invention and the total content of this active component is with IGD carbon-13 nmr spectra and the coupling of analysis quantitative means by the coupling formula.Be compared with existing technology, the present invention adopts IGD ¹³C NMR coupling finger-print has following several characteristics:

1. stability (repeatability): IGD ¹³The chemical shift data that C NMR obtains is second behind the decimal point, and explanation property is good, good reproducibility; The non-chromatographic condition of HPLC, GC (such as chromatographic column internal diameter, length, fixing mutually each component ratio, column temperature, sample size, detector sensitivity etc. of the phase trade mark, carrier granularity, flow rate of mobile phase, mixed flow) changes etc., the retention time data variation that obtains is very large, mean the variation of monolithic chromatogram figure, repeatability is bad.

2. globality (comprehensive): IGD ¹³The corresponding spectrum peak that comprises each the active component carbon in the sample in the C NMR finger-print; There are not this relation in HPLC, GC, UV, IR, MS.

3. reliability (unicity): IGD ¹³Carbon in C NMR spectrum peak and the sample on different activities composition and the different group thereof is strict one-to-one relationship; There are not this relation in HPLC, GC, UV, IR, MS.

4. feasibility (the easily property distinguished): IGD ¹³C NMR finger-print regularity is very strong, generally speaking, can belong to each the carbon peak in the collection of illustrative plates; HPLC, GC need reference substance; IR is difficult for resolving; The UV information content is few; MS then has the problems such as degree of ionization and matrix interference.

The present invention is directed to diversity, the complexity of active component in the Celastrus angulatus mother liquor, and efficient liquid-phase chromatograph finger print atlas and proton nmr spectra ( ¹H NMR) limitation of finger-print, make up IGD carbon-13 nmr spectra coupling fingerprint pattern technology, reflect and contain which beta-dihydroagarofuran sesquiterpene polyolester compound in the Celastrus angulatus mother liquor, and their content, proportionate relationship and total contents separately, reach the purpose to Celastrus angulatus mother liquor and aqueous emulsion quality testing and control.Stability and veracity, repeatability and feasibility compared with prior art improve a lot.So that Celastrus angulatus mother liquor and Celastrus angulatus aqueous emulsion definite ingredients, content are clear, quality controllable, stable performance, systematization and the standardization of the inherent composition Study of Celastrus angulatus mother liquor and aqueous emulsion thereof have been strengthened.

Celastrus angulatus mother liquor of the present invention (i.e. 10% Celastrus angulatus mother liquor) is compared with existing Celastrus angulatus mother liquor (i.e. 6% Celastrus angulatus mother liquor), and main advantage is embodied in the significantly raising of Celastrus angulatus content.

And, Celastrus angulatus aqueous emulsion (2%) by Celastrus angulatus mother liquor of the present invention (10%) preparation belongs to the friendly type agricultural chemicals of production environment, and 1% Celastrus angulatus missible oil of 6% Celastrus angulatus mother liquor preparation, owing to polluting larger, the feature of environmental protection is poor, the Ministry of Industry and Information Technology's the 4th explicit stipulation in former [2009] No. 29 files of worker: " from 1 day August in 2009, no longer issuing emulsifiable concentrates for agricultural chemicals product certificate of approval ", that is to say, its production is halted with other emulsifiable concentrates for agricultural chemicals products.

Microemulsion once caused people's worry because of emulsifier, cosolvent large usage quantity in the early stage of development, and less stable, cost are higher, microemulsion be not in the world formulation development trend [Hua Naizhen. the exploitation of water base chemical preparation and prospect. agricultural chemicals 2006,45 (12): 805.].In addition, 0.5% Celangulins ME no longer production because interim lot number expires.

Therefore, Celastrus angulatus mother liquor active component content of the present invention is high, and the control efficiency of Celastrus angulatus aqueous emulsion has surpassed 1% Celastrus angulatus missible oil, and its advantages shows that pollution is little, belongs to the friendly type agricultural chemicals of production environment; And instant effect, good stability, cost are low, are beneficial to popularization, have very high using value and economic worth.

Description of drawings

Fig. 1-a is embodiment 210% Celastrus angulatus mother liquor A(Xichuan County, henan Province) feature extraction thing IGD carbon-13 nmr spectra finger-print.

Fig. 1-b is embodiment 210% Celastrus angulatus mother liquor A(Xichuan County, henan Province) part, feature extraction thing IGD carbon-13 nmr spectra Fingerprints peak widens enlarged drawing.

Fig. 2-a is embodiment 210% Celastrus angulatus mother liquor B(Xichuan County, henan Province) feature extraction thing IGD carbon-13 nmr spectra finger-print.

Fig. 2-b-1 is embodiment 210% Celastrus angulatus mother liquor B(Xichuan County, henan Province) feature extraction thing IGD carbon-13 nmr spectra finger-print C-8 characteristic peak part widens enlarged drawing.

Fig. 2-b-2 is embodiment 210% Celastrus angulatus mother liquor B(Xichuan County, henan Province) feature extraction thing IGD carbon-13 nmr spectra finger-print C-15 characteristic peak part widens enlarged drawing.

Fig. 3-a is embodiment 210% Celastrus angulatus mother solution C (Xichuan County, henan Province) feature extraction thing IGD carbon-13 nmr spectra finger-print.

Fig. 3-b-1 is that embodiment 210% Celastrus angulatus mother solution C (Xichuan County, henan Province) feature extraction thing IGD carbon-13 nmr spectra finger-print C-8 characteristic peak part widens enlarged drawing.

Fig. 3-b-2 is that embodiment 210% Celastrus angulatus mother solution C (Xichuan County, henan Province) feature extraction thing IGD carbon-13 nmr spectra finger-print C-15 characteristic peak part widens enlarged drawing.

Fig. 4-a is embodiment 310% Celastrus angulatus mother liquor D(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print.

Fig. 4-b-1 is embodiment 310% Celastrus angulatus mother liquor D(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print C-8 characteristic peak part widens enlarged drawing.

Fig. 4-b-2 is embodiment 310% Celastrus angulatus mother liquor D(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print C-15 characteristic peak part widens enlarged drawing.

Fig. 5-a is embodiment 310% Celastrus angulatus mother liquor E(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print.

Fig. 5-b-1 is embodiment 310% Celastrus angulatus mother liquor E(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print C-8 characteristic peak part widens enlarged drawing.

Fig. 5-b-2 is embodiment 310% Celastrus angulatus mother liquor E(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print C-15 characteristic peak part widens enlarged drawing.

Fig. 6-a is embodiment 310% Celastrus angulatus mother liquor F(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print.

Fig. 6-b-1 is embodiment 310% Celastrus angulatus mother liquor F(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print C-8 characteristic peak part widens enlarged drawing.

Fig. 6-b-2 is embodiment 310% Celastrus angulatus mother liquor F(Baoji) feature extraction thing IGD carbon-13 nmr spectra finger-print C-15 characteristic peak part widens enlarged drawing.

Fig. 7-a is embodiment 56% Celastrus angulatus mother liquor feature extraction thing IGD carbon-13 nmr spectra finger-print.

Fig. 7-b-1 is that embodiment 56% Celastrus angulatus mother liquor feature extraction thing IGD carbon-13 nmr spectra finger-print C-8 characteristic peak part widens enlarged drawing.

Fig. 7-b-2 is that embodiment 56% Celastrus angulatus mother liquor feature extraction thing IGD carbon-13 nmr spectra finger-print C-15 characteristic peak (containing C-15) part widens enlarged drawing.

Fig. 8 is that 10% Celastrus angulatus mother liquor feature extraction thing obtains the degree flow chart.

Embodiment

Below in conjunction with specific embodiment technical scheme of the present invention is described in further detail, but protection scope of the present invention is not limited to this.

The preparation method of 1.10% Celastrus angulatus mother liquor, aqueous emulsion

(1) 10% Celastrus angulatus mother liquor

Take by weighing the Celastrus angulatus root that dries in the shade or/and skin, crossing 10 ~ 24 mesh sieves after pulverizing, is that the benzene of 5 ~ 7 times of amounts extracts 1 ~ 2 time at 45 ~ 55 ℃ of lower ultrasonic circulations with volume, extracts 35 ~ 55 minutes at every turn, be evaporated to density after filtrate merges and be about 1.1 ~ 1.3 medicinal extract, namely get 10% Celastrus angulatus mother liquor.

(2) 2% Celastrus angulatus aqueous emulsions

After 10% Celastrus angulatus mother liquor and emulsifier mix by a certain percentage, add certain proportion antifreeze and water and supply surplus, shear homogeneous after, namely get 2% Celastrus angulatus aqueous emulsion.

2% Celastrus angulatus aqueous emulsion mixed proportion is as follows:

10% Celastrus angulatus mother liquor 15 ~ 20%, emulsifier 8 ~ 12%, antifreeze 3 ~ 4%, water are supplied surplus.The percentage sign here " % " is mass percent.Wherein, emulsifier alkylaryl polyoxyethylene poly-oxygen propylene aether is available from Shandong natural laws biotechnology Co., Ltd.

(3) instrument, reagent and material

Ultrasonic circulation extracts circulator DST-100 type, Shanghai whetstone real machine tool equipment Co., Ltd.

2.IGD carbon-13 nmr spectra finger print quality detecting method

(1) IGD carbon-13 nmr spectra finger print quality detecting method research step

1) the feature extraction thing obtains program research

Accurately take by weighing 10% Celastrus angulatus mother liquor, filter rear at 55 ~ 65 ℃ of lower reduced pressure concentrations, reclaim the solvent evaporate to dryness to the medicinal extract shape, medicinal extract with 6 ~ 10 times of amount volumes, the dissolving of 80% ~ 90% methyl alcohol after, use again the petroleum ether extraction 1 ~ 2 time of 3 ~ 5 times of amount volumes, reduced pressure concentration after 80% ~ 90% methanol layer filters reclaims solvent to doing, and namely gets Celastrus angulatus mother liquor feature extraction thing.Its program flow diagram as shown in Figure 8, T10 refers to contain 10 kinds of active ingredient substances.

2) feature extraction thing IGD carbon-13 nmr spectra finger-print detects

Get 10% Celastrus angulatus mother liquor feature extraction thing 80mg, be dissolved in 0.5mLCDCl ₃In, make IGD carbon-13 nmr spectra finger-print and detect, namely obtain IGD carbon-13 nmr spectra finger-print.

3) feature extraction thing IGD carbon-13 nmr spectra finger-print is resolved

1. differentiate

In the feature extraction thing IGD carbon-13 nmr spectra finger-print, should clearly illustrate the characteristic signal of 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound, and contain Celangulin A, E, H, celangulatin C; Celangulin B, C, F, G, Celastrus angulatus II, Celastrus angulatus IV signal.

Concrete data are as follows:

δ _C74.9-75.8 or 70.5-70.9,66.9-68.1,41.1-42.7,72.1-72.5,91.3-91.7 are respectively A

ring

1,2,3,4,5 alicyclic ring carbon signals; 76.2-77.0,53.0-53.8,73.3-74.7 or 75.6-76.7,74.9-75.5 or 70.6-72.6,50.1-50.8 or 53.9-54.5 is respectively B ring 6,7,8,9,10 alicyclic ring carbon signals, 82.5 ~ 84.6 is 4-OH-beta-dihydroagarofuran sesquiterpene polyolester Compound C-11 carbon signal, 29.3-30.1,26.2-26.4 or 25.4-25.7,24.1-24.6 is 4-OH-beta-dihydroagarofuran sesquiterpene polyolester Compound C-12, C-13, C-14 methyl carbon signal, 61.4-61.8 or 65.0-65.7 are 4-OH-beta-dihydroagarofuran sesquiterpene polyolester Compound C-15 mesomethylene carbon signal.

2. each active component characteristic peak is chosen in the 10% Celastrus angulatus mother liquor feature extraction thing

Owing to containing a series of active component 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compounds in the feature extraction thing, the carbon peak intersects severe, in order to measure the ratio of each active component, must select the larger respective peaks of chemical shift difference as characteristic peak., investigate through reality for this reason, selected δ _C60.0 one group of C-15 peak and δ about ~ 66.0 _C73.8 Celangulin A C-8 peak, the left and right sides.Its former because: generally speaking, the C-15 peak is easy to identification as connecting oxygen carbon; The C-15 position is subjected to the γ effect of the upper α of C-9 and β bit substituent different, and its chemical shift difference is larger.Chemical shift such as Celangulin A, E, H, celangulatin C is δ _C61.0 about; The chemical shift of Celangulin B, C, F, G, Celastrus angulatus II, Celastrus angulatus IV is δ _C65.0 about.Because the C-15 position overlap of peaks of Celangulin A, E, celangulatin C and Celangulin C, F, Celastrus angulatus II, alternative is selected δ _C73.3-74.7 it is overlapping that C-8 peak, the left and right sides does not have as characteristic peak difference Celangulin A, Celangulin E, celangulatin C peak, selects δ _C82.5-83.9 C-11 peak, the left and right sides is as characteristic peak difference Celangulin C, F, Celastrus angulatus II.

3. the selection of criterion referenced product

Celangulin A is one of main active insecticidal components of Insecticidal Plant Celastrus Angulatus, and standard items are easy to get.Therefore, select Celangulin A as the criterion referenced product.

4) adopt HPLC to measure the content of Celangulin A in 10% Celastrus angulatus mother liquor or the 2% Celastrus angulatus aqueous emulsion

1. HPLC detects

I) chromatographic condition

Instrument: the Agilent(Agilent) 1200

Mobile phase: acetonitrile: water=55:45

Flow velocity: 1mL/min

Chromatographic column: Thermo C ₁₈(Thermo octadecyl silane) 250*4.6mm

Detector: ultraviolet

Wavelength: 232nm

Sample size: 20 μ L;

Ii) preparation of criterion referenced product solution

Accurately take by weighing Celangulin A5mg, put in the 50mL volumetric flask, to scale, namely get criterion referenced product solution (Celangulin A100 μ g/mL) with the methyl alcohol dissolved dilution after shaking up.

Iii) calibration curve and detection limit

Concentration range: 1 ~ 100 μ g/mL(ppm); Criterion referenced product concentration is respectively: 1 μ g/mL, 5 μ g/mL, 50 μ g/mL, 100 μ g/mL.

Under above-mentioned chromatographic condition, carry out HPLC and analyze, Celangulin A total peak area Y to the equation of linear regression of concentration C is:

Y=23.85113*C+1.25004（n=5,R=0.9995）。

Detect and be limited to: 0.5ug/mL (S/N=3).

According to canonical plotting, in selected concentration range, the working curve linear relation of the standard liquid of Celangulin A is good.

Iv) preparation of need testing solution

Accurately take by weighing 10% Celastrus angulatus mother liquor or 2% Celastrus angulatus aqueous emulsion 200mg in the 100mL volumetric flask, add an amount of methyl alcohol dissolving, be diluted to scale behind the sonic oscillation, namely get 10% Celastrus angulatus mother liquor or 2% Celastrus angulatus aqueous emulsion need testing solution after shaking up.

V) precision is measured

Need testing solution repeats sample introduction 3 times, peak area relative standard deviation RSD=0.22%, retention time relative standard deviation RSD=0.11%.

Vi) mensuration of test sample

Draw each need testing solution, sample introduction is surveyed its peak area, tries to achieve Celangulin A content.

Vii) determination of recovery rates

Employing standard adding method, mark-on 100 μ g/mL in test sample, average recovery rate is between 89.0 ~ 101.2%.

2. Celangulin A absolute content calculates

I) calculate Celangulin A mass concentration in the need testing solution by following formula

C_{X} = C_{R} \times \frac{A_{X}}{A_{R}}

C _X: Celangulin A mass concentration (ug/mL) in 10% Celastrus angulatus mother liquor or the 2% Celastrus angulatus aqueous emulsion need testing solution;

C _R: criterion referenced product solution Celangulin A mass concentration (ug/mL);

A _X: the peak area of Celangulin A in the 10% Celastrus angulatus mother liquor of being measured by HPLC or the 2% Celastrus angulatus aqueous emulsion need testing solution;

A _R: by the peak area of Celangulin A in the criterion referenced product solution of HPLC mensuration.

Ii) calculate 10% Celastrus angulatus mother liquor or 2% Celastrus angulatus aqueous emulsion Celangulin A quality percentage composition by following formula

X ₁(%): Celangulin A quality percentage composition in 10% Celastrus angulatus mother liquor or the 2% Celastrus angulatus aqueous emulsion, the i.e. absolute content of criterion referenced product Celangulin A (quality percentage composition);

W _{Test sample}: the 10% Celastrus angulatus mother liquor that takes by weighing or 2% Celastrus angulatus aqueous emulsion quality (mg).

5) calculate main active content in the 10% Celastrus angulatus mother liquor by the coupling formula

X_{n} = \frac{X_{1} M_{n} h_{n}}{M_{1} h_{1}}

X _n(%): a certain active component quality percentage composition % in the 10% Celastrus angulatus mother liquor;

X ₁(%): Celangulin A quality percentage composition %, the i.e. absolute content of criterion referenced product Celangulin A (quality percentage composition) in the 10% Celastrus angulatus mother liquor;

M ₁: Celangulin A(criterion referenced product) molecular weight;

h ₁: by Celangulin A(criterion referenced product in the 10% Celastrus angulatus mother liquor feature extraction thing of IGD carbon-13 nmr spectra determining fingerprint pattern) characteristic peak peak intensity (peak height);

M _n: a certain active component molecular weight in the 10% Celastrus angulatus mother liquor feature extraction thing;

h _n: by a certain active component characteristic peak peak intensity (peak height) in the 10% Celastrus angulatus mother liquor feature extraction thing of IGD carbon-13 nmr spectra determining fingerprint pattern.

6) calculate main single active component coefficient and overall coefficient in the 10% Celastrus angulatus mother liquor by the coupling formula

Coefficient formulas

F_{n} = \frac{M_{n} h_{n}}{M_{1} h_{1}}

F _n: a certain active component and Celangulin A(criterion referenced product in the 10% Celastrus angulatus mother liquor) ratio coefficient of quality percentage composition;

M ₁: Celangulin A(criterion referenced product) molecular weight;

Overall coefficient is F _nAdd and.

This coefficient F _nBe applicable to too calculate active component and active component group in the 2% Celastrus angulatus aqueous emulsion.

7) main active content and calculation of total in the 2% Celastrus angulatus aqueous emulsion

W _n（%）=W ₁（%）F _n

W _n: a certain active component quality percentage composition % in the 2% Celastrus angulatus aqueous emulsion;

W ₁: Celangulin A quality percentage composition %, the i.e. absolute content of criterion referenced product Celangulin A in the 2% Celastrus angulatus aqueous emulsion.

(2) instrument, reagent and material

Nuclear magnetic resonance chemical analyser Bruker DPX400 type.

Mass spectrograph: Waters Micromass Q-Tof MicroTM 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 separatory funnel.

DE-52AA Rotary Evaporators: Shanghai Yarong Biochemical Instrument Plant.

DEF-6020 type vacuum drying chamber: the upper grand experimental facilities of Nereid Co., Ltd.

Column chromatography silica gel G and tlc silica gel H: Haiyang Chemical Plant, Qingdao.

Silica gel column chromatography 6cm * 70cm(diameter * highly).

Celastrus angulatus medicinal material (Xichuan County, henan Province, in November, 2009, a large amount of purchases were from local), Celastrus angulatus medicinal material (Baoji, a large amount of purchases were from local in 2009 11) are all identified through professor Zhu Changshan of In Henan Agriculture university.

Celangulin A, criterion referenced product, laboratory self-control (identifying through spectroscopic data).

Reagent: chromatographically pure (methyl alcohol, Tianjin Siyou Fine Chemicals Co., Ltd.) and analyze pure (Tianjin Chemical Reagents Factory No.1).

(3) basic research

1) separation and Extraction flow process

Take by weighing the Xichuan County, henan Province that dries in the shade and produce Celastrus angulatus root, skin 1kg, pulverize, measure volume benzene refluxing extraction 3 times with 6 times, filtrate merges rear 60 ℃ of reduced pressure concentrations, reclaim solvent to the medicinal extract shape, after the methyl alcohol dissolving of medicinal extract with 6 times of amount bulk purity 80%, the petroleum ether extraction of 60mL 1 time, reduced pressure concentration after methanol layer filters reclaims solvent and gets medicinal extract (27.5g).Get this medicinal extract and separate with silica gel column chromatography, (10:1 ~ 4:6) dicyandiamide solution carries out gradient elution, and every 250mL collects 1 part, merges identical cut with petroleum ether-ethyl acetate.The 38th part through the preparative chromatography purifying, gets Celangulin E(18mg); The 42nd ~ 43 part gets Celangulin A sterling (60mg); The 47th part through the preparation purifying get celangulatin C (41mg); The 54th part through the preparative chromatography purifying, gets Celangulin B(25mg), Celangulin H(40mg); The 61st part gets Celangulin G(95mg); The 69th ~ 70 part through the preparative chromatography purifying, gets Celangulin F(45mg).

Take by weighing the Henan Xixia Mine root bark of celastrus angulatus powder 1kg that dries in the shade, add under 3 times of amount benzene 80 ℃ of temperature refluxing extraction three times, filter merging filtrate.Reduced vacuum simmer down to medicinal extract.According to said method extract altogether root bark of celastrus angulatus powder 7kg, merge all crude extracts.Crude extract is through reduced vacuum simmer down to pasty masses (crude extract).Crude extract medicinal extract is 206g.Add the dissolving of 4200mL80% methyl alcohol in crude extract medicinal extract after, 300mL petroleum ether extraction 1 time gets methyl alcohol part medicinal extract 113.3g.Get this medicinal extract and separate with silica gel column chromatography, (10:1 ~ 4:6) dicyandiamide solution carries out gradient elution (every 500mL collects a), prepares purifying through high efficiency liquid phase and obtains following β-dihydroagarofuran polyol ester compounds with petroleum ether-ethyl acetate.The 66th ~ 70 part through the preparation purifying, get Celangulin A sterling (60mg); The 80th part through the preparation purifying get Celangulin E(18mg); The 127th part through the preparation purifying get Celangulin H(40mg) and Celangulin B(60mg); The 161st part through the preparation purifying get Celangulin G(95mg), the 203rd part through the preparation purifying get Celangulin C.

Celastrus angulatus II, IV data see document [Wu Wenjun. vegetable insecticide Celastrus angulatus research with use. Chemical Industry Press, 2010].

2) structure and the carbon-13 nmr spectra data of main active in 10% Celastrus angulatus mother liquor and 2% aqueous emulsion

Celangulin A:R ₁=R ₄=OiBu, R ₂=R ₅=H, R ₃=OBz, R ₆=OH, R ₇=OAc

Celangulin B:R ₁=R ₅=OiBu, R ₂=OFu, R ₃=R ₄=H, R ₆=R ₇=OAc

Celangulin C:R ₁=R ₅=OiBu, R ₂=OBu, R ₃=R ₄=H, R ₆=R ₇=OAc

Celangulin E:R ₁=OiPet, R ₂=R ₅=H, R ₃=OBz, R ₄=OiBu, R ₆=OH, R ₇=OAc

Celangulin F:R ₁=R ₆=R ₇=OAc, R ₂=R ₅=OFu, R ₃=R ₄=H

Celangulin G:R ₁=R ₆=R ₇=OAc, R ₂=OFu, R ₃=R ₄=H, R ₅=OiBu

Celangulin H:R ₁=R ₇=OAc, R ₂=R ₅=H, R ₃=OBz, R ₄=OiBu, R ₆=OH

Celangulatin C:R ₁=OiBu,R ₂=R ₅=H,R ₃=OBz,R ₄=R ₇=OAc,R ₆=OH

Celastrus angulatus II:R ₁=OiBu, R ₂=OBz, R ₃=R ₄=H, R ₅=R ₆=R ₇=OAc

Celastrus angulatus IV:R ₁=OiBu, R ₂=R ₅=OFu, R ₃=R ₄=H, R ₆=R ₇=OAc

Celangulin A

¹³C NMR(100MHz,CDCl ₃)δ _C:75.01(C-1)，67.26(C-2),41.15(C-3),72.13(C-4)，91.41(C-5)，76.89(C-6)，53.51(C-7)，73.69(C-8)，75.28(C-9)，50.52(C-10),84.54(C-11),30.05(C-12)，26.35(C-13)，24.14(C-14)，61.69(C-15)

OAC:169.49,169.60，20.50，21.13

OiBu:175.82,176.77,34.09,34.31,18.46,18.66,19.06,19.14

OBz:165.67,129.26,129.44,128.63,133.45

Celangulin B

¹³C NMR(100MHz,CDCl ₃)δ _C:70.63(C-1)，67.96(C-2),42.00(C-3),69.84(C-4)，91.33(C-5)，75.39(C-6)，52.97(C-7)，76.05(C-8)，71.44(C-9)，53.87(C-10),83.44(C-11),29.59(C-12)，25.45(C-13)，24.47(C-14)，65.45(C-15)

OAC:169.54,169.66,169.79,20.55,21.12,21.48

OiBu:175.74,176.90,33.95,34.10,18.73,18.99,19.00,19.06

OFu:160.91,148.99,117.81,109.69,144.00

Celangulin C

¹³C NMR(100MHz,CDCl ₃)δ _C:70.86(C-1),67.98(C-2),42.04(C-3),69.88(C-4)，91.42(C-5),75.48(C-6),53.04(C-7),76.19(C-8),72.03(C-9),54.10(C-10),83.54(C-11),29.61(C-12),25.66(C-13),24.50(C-14),65.55(C-15);

OA _C: 169.44,169.57,169.26,20.38,21.13*, 21.50* (CH ₃) (the * ownership is interchangeable)

OiBu:175.77,176.95,33.98,34.12,18.78,18.91,19.04,19.08

OBz:164.52,128.31,130.18,128.47,133.87

Celangulin E

¹³C NMR(100MHz,CDCl ₃)δ _C:75.09(C-1)，67.31(C-2),41.21(C-3),72.15(C-4)，91.49(C-5)，76.92(C-6)，53.53(C-7)，73.80(C-8)，75.23(C-9)，50.49(C-10),84.54(C-11),30.08(C-12)，26.31(C-13)，24.06(C-14)，61.68(C-15)

OAC:169.55,169.46,20.48,21.13

OiBu:175.72,34.11,18.48,18.64

OiPet:176.19,41.56,26.59,11.69,16.92

OBz:165.68,129.36,129.49,128.61,133.42

Celangulin F

¹³C NMR(100MHz,CDCl ₃)δ _C:70.53(C-1)，67.80(C-2),41.93(C-3),69.90(C-4)，91.31(C-5)，75.34(C-6)，53.80(C-7)，76.54(C-8)，71.65(C-9)，54.33(C-10),83.12(C-11),25.55(C-12)，29.51(C-13)，24.32(C-14)，65.56(C-15)

OAC:169.51,169.75,169.86,170.52,20.47,21.13,21.52,21.11

OFu:161.61,160.53,148.69,148.87,118.84,118.05,109.96,109.78,144.01,144.01

Celangulin G

¹³C NMR(100MHz,CDCl ₃)δ _C:70.49(C-1)，67.82(C-2),41.92(C-3),69.81(C-4)，91.51(C-5)，75.00(C-6)，53.34(C-7)，76.10(C-8)，72.65(C-9)，53.97(C-10),83.27(C-11),29.45(C-123)，25.48(C-13)，24.35(C-14)，65.67(C-15)

OAC:169.47,169.69,169.74,,170.53,20.45,21.14,21.52,21.04

OiBu:175.86,33.92,18.81,18.89

OFu:160.91,148.99,117.83,109.71,144.02

Celangulin H

¹³C NMR(100MHz,CDCl ₃)δ _C:74.89(C-1)，67.31(C-2),41.10(C-3),72.11(C-4)，91.44(C-5)，76.87(C-6)，53.59(C-7)，73.36(C-8)，75.40(C-9)，50.64(C-10),84.52(C-11),30.03(C-123)，26.24(C-132)，24.23(C-14)，61.38(C-15)

OAC:169.39,169.55,170.28,20.49,21.10,21.47

OiBu:175.88,34.02,18.41,18.58

OBz:165.72,129.23,129.40,128.64,133.40

Celangulatin C(Celastrus angulatus C)

¹³C NMR(100MHz,CDCl ₃)：δ _C：75.07（C-1），67.31（C-2），41.19（C-3），72.14（C-4），91.45（C-5），76.90（C-6），53.48（C-7），74.17（C-8），75.34（C-9），50.64（C-10），84.58（C-11），30.05（C-12），26.28（C-13），24.20（C-14），61.73（C-15）

OA _C:169.44,169.54,169.95,20.44,21.10,20.82

OiBu：176.72，34.34，19.05，19.12

OBz：165.63，129.49，129.28,129.66,133.47

Celastrus angulatus II

¹³C NMR (100MHz, CDCl ₃) δ _C: 68.0* (C-1), 70.8* (C-2), 42.7 (C-3), 69.9 (C-4), 91.5 (C-5), 72.5* (C-6), 53.2 (C-7), 75.8* (C-8), 76.7* (C-9), 54.4 (C-10), 83.3 (C-11), 29.5 (C-12), 24.6 (C-13), 24.6C-14), 65.6 (C-15) (the * ownership is interchangeable)

OAC:169.3,169.2×2,21.0×2,21.4

OiBu:176.9,34.1,19.0,19.1

OBz：164.5，129.5，130.2,128.4,133.8

Celastrus angulatus IV

¹³C NMR (100MHz, CDCl ₃) δ _C: 70.3* (C-1), 70.6* (C-2), 42.0 (C-3), 69.9 (C-4), 91.4 (C-5), 75.6* (C-6), 53.6 (C-7), 76.1* (C-8), 68.0* (C-9), 54.4 (C-10), 83.4 (C-11), 29.6 (C-12), 25.5 (C-13), 24.4 (C-14), 65.0 (C-15) (the * ownership is interchangeable)

OAC:169.6×3,20.5,21.0,21.4

OiBu:176.7,34.0,18.8,18.9

OFu:161.3,160.6,148.9,148.7,118.8,118.1,109.8,109.7,143.9×2

Preparation and the preventive effect of embodiment 1:10% Celastrus angulatus mother liquor and 2% Celastrus angulatus aqueous emulsion

The preparation of (1) 10% Celastrus angulatus mother liquor

Celastrus angulatus root and the skin of (24 order) up to specification after will sieving, be the benzene (raw material: benzene=kg:L of 5 ~ 7 times of amounts according to volume, being the Celastrus angulatus root and the benzene of skin with 5 ~ 7L of 1kg) 50 ℃ of ultrasonic circulations extract 35 ~ 55min (ultrasonic power 500W) 1 ~ 2 time (each extraction conditions is all consistent), reduced pressure concentration became 1.1 ~ 1.3 medicinal extract after filtrate merged, and namely obtained 10% Celastrus angulatus mother liquor.Concrete data see the following form.

The preparation of (2) 2% Celastrus angulatus aqueous emulsions

Place reactor with detecting 10% qualified Celastrus angulatus mother liquor, the auxiliary material (emulsifier and antifreeze) according in the proportional quantity adding following table stirs, and solubilizer is stirred to fully dissolving to stipulating content again, obtains 2% Celastrus angulatus aqueous emulsion.Configuration proportion sees the following form.

(3) field trial

The experiment of 2% Celastrus angulatus aqueous emulsion field efficacy is given birth to the pesticide field efficacy medicine test criterion of surveying the chamber formulation with reference to the Institute for the Control of Agrochemicals of the Ministry of Agriculture,PRC and is carried out.

1) materials and methods

A. reagent agent

2% Celastrus angulatus aqueous emulsion (formulated by above-mentioned 10% Celastrus angulatus mother liquor A), 1% Celastrus angulatus missible oil (production of Xinxiang City east wind responsibility Co., Ltd).

B. controlling object and trial crops

Controlling object is cabbage caterpillar (pieris rapae).Trial crops is wild cabbage.

C. experimental field reach crop

Experimental field be located at Yuxi Tonghai County, Yunnan Province Experimental Base, black sandy loam soil property, fertility is even, and managerial skills are consistent, and the plantation wild cabbage is brassicaceous vegetable, and cabbage caterpillar all has generation over the years.

D. chemicals treatment

If 2% Celastrus angulatus aqueous emulsion 800,3 processing of 1000,1200 times of liquid are diluted 800 times of liquid as the contrast medicament take 1% Celastrus angulatus missible oil to water, the spray clear water is blank.3 repetitions are established in every processing, totally 18 residential quarters.Residential quarter area 20m ², random district group is arranged.

E. spraying time and method

Be administered to for the first time and carried out on October 13rd, 2011, cause harm the Sheng phase on the occasion of the cabbage caterpillar larva.Testing the same day is partly cloudy weather, 25.5 ℃ of mean temperature of air.With the dispenser of bodyguard's board WS-16P knapsack hand sprayer.Medicament is diluted to the test desired concn, is sprayed onto that the blade face is moistening is advisable without dropping liquid.

F. control time and method

Decide 5 points in every residential quarter with 5 point samplings before the dispenser, decide 4 of wild cabbages at every, listing mark, and the radix of investigation cabbage caterpillar.Behind the spray medicine 3,5,10d is at the remaining borer population alive of the wild cabbage of mark investigation, calculate each treatment region control efficiency and carry out significance of difference test.

Preventive effect (%)=[the mite number of living before the borer population * check plot medicine of living behind the 1 one treatment region medicines/(borer population of living before the borer population * treatment region medicine of living behind the medicine of check plot)] * 100

2) statistic analysis result

Cabbage caterpillar field efficacy experiment analysis results sees the following form.Can find out from table, it is better that 2% Celastrus angulatus aqueous emulsion is diluted to 800 ~ 1000 times of preventive effect effects, and 3 ~ 5 days control efficiency is 80.18% ~ 91.86% behind its medicine; Best to 800 times for the treatment of region preventive effects of water dilution, 3 ~ 5 days control efficiency is 85.61% ~ 91.86% behind its medicine; Obviously be better than contrasting medicament 1% Celastrus angulatus missible oil to 800 times of preventive effects of water dilution; 10 days its lasting periods.

According to other aqueous emulsions of the present embodiment preparation with similar by the aqueous emulsion effect of 10% Celastrus angulatus mother liquor A preparation, take by 10% Celastrus angulatus mother liquor A preparation for optimum.

Embodiment 2:10% Celastrus angulatus mother liquor (Xichuan County, henan Province) IGD carbon-13 nmr spectra coupling finger print quality detecting method

(1) mother liquor feature extraction thing preparation

Select 10% Celastrus angulatus mother liquor (Xichuan County, henan Province), prepare as follows 10% Celastrus angulatus mother liquor feature extraction thing: take by weighing 10% Celastrus angulatus mother liquor 20g, filter reduced pressure concentration under rear 60 ℃ of temperature, reclaim solvent to the medicinal extract shape, medicinal extract is after 80% the methyl alcohol dissolving with 6 times of amounts volume (120mL), mass ratioes, the petroleum ether extraction of 3 times of amount volumes (60mL) 1 time, each extraction 30min, reduced pressure concentration after methanol layer filters, reclaim solvent to doing, namely get 10% Celastrus angulatus mother liquor (Xichuan County, henan Province) feature extraction thing.

(2) feature extraction thing IGD carbon-13 nmr spectra test

Get 10% Celastrus angulatus mother liquor (Xichuan County, henan Province) feature extraction thing 80mg, be dissolved in 0.5mLCDCl ₃In, make the IGD carbon-13 nmr spectra.

(3) IGD carbon-13 nmr spectra finger-print is resolved

1) differentiates

In the IGD carbon-13 nmr spectra finger-print of 10% Celastrus angulatus mother liquor (Xichuan County, henan Province) feature extraction thing, clearly illustrate the characteristic signal of 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound.According to the chemical shift of its parent nucleus signal and C-15 characteristic peak signal thereof, proving conclusively 10 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound Celangulin A, B, C, E, F, G, H, celangulatin C, Celastrus angulatus II, Celastrus angulatus IV all has corresponding NMR signal in IGD carbon-13 nmr spectra finger-print.IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 1-a, Fig. 2-a and Fig. 3-a.

2) 10% Celastrus angulatus mother liquor A-C(Xichuan County, henan Province) each active component ratio measuring result is as follows:

1. 10% Celastrus angulatus mother liquor A(Xichuan County, henan Province)

IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 1-a, and its characteristic peak part widens enlarged drawing and sees accompanying drawing 1-b.

2. 10% Celastrus angulatus mother liquor B(Xichuan County, henan Province)

IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 2-a, and its characteristic peak layout widens enlarged drawing and sees accompanying drawing 2-b-1 and 2-b-2.

3. 10% Celastrus angulatus mother solution C (Xichuan County, henan Province)

IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 3-a, and its characteristic peak part widens enlarged drawing and sees accompanying drawing 3-b-1 and 3-b-2.

(4) 10% Celastrus angulatus mother liquor A-C(Xichuan County, henan Provinces) result is as follows for Celangulin A concentration assay:

	Mother liquor A	Mother liquor B	Mother solution C
				Celangulin A quality percentage composition in the mother liquor	2.85%	2.87%	3.03%

(5) 10% Celastrus angulatus mother liquor A-C(Xichuan County, henan Provinces) each content determinations of active component result is as follows in:

1. Celastrus angulatus mother liquor A(Xichuan County, henan Province)

The molecular formula of Celangulin C+ Celastrus angulatus II+ Celangulin F is with the molecular formula of Celangulin C as representative in the following table, the molecular formula of Celangulin A+ Celangulin E+celangulatin C is with the molecular formula of Celangulin A as representative, and the molecular formula of Celastrus angulatus II+ Celangulin F is with the molecular formula (following examples all with embodiment 1) of Celangulin II as representative.

2. 10% Celastrus angulatus mother liquor B(Xichuan County, henan Province)

3. 10% Celastrus angulatus mother solution C (Xichuan County, henan Province)

Embodiment 3:10% Celastrus angulatus mother liquor (Baoji) IGD carbon-13 nmr spectra coupling finger print quality detecting method

(1) mother liquor feature extraction thing preparation

Difference from Example 2 is to select the mother liquor of Baoji, different place of production medicinal material preparation, and with the methyl alcohol dissolving of 10 times of amount volumes, mass ratio 90%, the petroleum ether extraction of 5 times of amount volumes (100mL) 1 time, other are identical.

(2) mother liquor feature extraction thing IGD carbon-13 nmr spectra test

Identical with embodiment 2.

(3) 10% Celastrus angulatus mother liquor (Baoji) IGD carbon-13 nmr spectra coupling finger-prints are resolved

1) differentiates

In the IGD carbon-13 nmr spectra finger-print of 10% Celastrus angulatus mother liquor (Baoji) feature extraction thing, clearly illustrate the characteristic signal of 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound.According to the chemical shift of its parent nucleus signal and C-15 characteristic peak signal thereof, proving conclusively 10 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound Celangulin A, B, C, E, F, G, H, celangulatin C, Celastrus angulatus II, Celastrus angulatus IV all has corresponding NMR signal in IGD carbon-13 nmr spectra finger-print.IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 4-a, Fig. 5-a and Fig. 6-a.

2) each active component ratio measuring result of 10% Celastrus angulatus mother liquor (Baoji) is as follows:

1. 10% Celastrus angulatus mother liquor D(Baoji)

IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 4-a, and its characteristic peak part widens enlarged drawing and sees accompanying drawing 4-b-1 and 4-b-2.

2. 10% Celastrus angulatus mother liquor E(Baoji)

IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 5-a, and its characteristic peak part widens enlarged drawing and sees accompanying drawing 5-b-1 and 5-b-2.

3. 10% Celastrus angulatus mother liquor F(Baoji)

IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 6-a, and its characteristic peak part widens enlarged drawing and sees accompanying drawing 6-b-1 and 6-b-2.

(4) 10% Celastrus angulatus mother liquor D-F(Baojis) result is as follows for Celangulin A concentration assay:

	A	B	C
				Celangulin A quality percentage composition in the mother liquor	2.16%	2.05%	1.96%

(5) 10% Celastrus angulatus mother liquor D-F(Baojis) the content determinations of active component result is as follows in:

1. 10% Celastrus angulatus mother liquor D(Baoji)

2. 10% Celastrus angulatus mother liquor E(Baoji)

3. 10% Celastrus angulatus mother liquor F(Baoji)

Each content determinations of active component in the embodiment 4:2% Celastrus angulatus aqueous emulsion

(1) 10% Celastrus angulatus mother liquor Celastrus angulatus coefficient calculations

Get the 10% Celastrus angulatus mother liquor A-C(Xichuan County, henan Province of embodiment 2) mean value of Celastrus angulatus overall coefficient is as Celastrus angulatus mother liquor (Xichuan County, henan Province) Celastrus angulatus overall coefficient.

Get the 10% Celastrus angulatus mother liquor D-F(Baoji of embodiment 3) mean value of Celastrus angulatus overall coefficient is as Celastrus angulatus mother liquor (Baoji) Celastrus angulatus overall coefficient.

Result of calculation is as follows:

The medicinal material place of production	Celastrus angulatus mother liquor Celastrus angulatus overall coefficient
		Xichuan County, henan Province	5.72
The Baoji	6.14

(2) 2% Celastrus angulatus aqueous emulsion Celangulin A concentration assay results are as follows:

(3) 2% Celastrus angulatus aqueous emulsion Celastrus angulatus cubages

Wherein, I ~ VI is different batches 2% Celastrus angulatus aqueous emulsion.

Embodiment 5:6% Celastrus angulatus mother liquor IGD carbon-13 nmr spectra coupling finger print quality detecting method

10% Celastrus angulatus mother liquor (Xichuan County, henan Province) IGD carbon-13 nmr spectra coupling finger print quality detecting method with reference to embodiment 2.

(1) mother liquor feature extraction thing preparation

Select 6% Celastrus angulatus mother liquor, prepare as follows 10% Celastrus angulatus mother liquor feature extraction thing: take by weighing 10% Celastrus angulatus mother liquor 20g, filter reduced pressure concentration under rear 60 ℃ of temperature, reclaim solvent to the medicinal extract shape, medicinal extract is after 80% the methyl alcohol dissolving with 6 times of amounts volume (120mL), mass ratioes, the petroleum ether extraction of 3 times of amount volumes (60mL) 1 time, reduced pressure concentration after methanol layer filters, reclaim solvent to doing, namely get 6% Celastrus angulatus mother liquor feature extraction thing.

(2) feature extraction thing IGD carbon-13 nmr spectra test

Get 6% Celastrus angulatus mother liquor feature extraction thing 80mg, be dissolved in 0.5mL CDCl ₃In, make the IGD carbon-13 nmr spectra.

(3) IGD carbon-13 nmr spectra finger-print is resolved

1) differentiates

In the IGD carbon-13 nmr spectra finger-print of 6% Celastrus angulatus mother liquor feature extraction thing, clearly illustrate the characteristic signal of 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound.According to the chemical shift of its parent nucleus signal and C-15 characteristic peak signal thereof, proving conclusively 10 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound Celangulin A, B, C, E, F, G, H, celangulatin C, Celastrus angulatus II, Celastrus angulatus IV all has corresponding NMR signal in IGD carbon-13 nmr spectra finger-print.

2) each active component ratio measuring result of 6% Celastrus angulatus mother liquor is as follows:

IGD carbon-13 nmr spectra finger-print is seen accompanying drawing 7-a, and its characteristic peak part widens enlarged drawing and sees accompanying drawing 7-b-1 and 7-b-2.

(4) 6% Celastrus angulatus mother liquor Celangulin A concentration assay results are as follows:

Celangulin A quality percentage composition in the 6% Celastrus angulatus mother liquor

1.75%

Each content determinations of active component result is as follows in (5) the 6% Celastrus angulatus mother liquors (Xichuan County, henan Province):

Embodiment 6:1% Celastrus angulatus missible oil IGD carbon-13 nmr spectra coupling finger print quality detecting method

2% Celastrus angulatus aqueous emulsion IGD carbon-13 nmr spectra coupling finger print quality detecting method with reference to embodiment 4.

(1) 6% Celastrus angulatus mother liquor Celastrus angulatus coefficient calculations

Get the 6% Celastrus angulatus mother liquor Celastrus angulatus overall coefficient of embodiment 5.

(2) 1% Celastrus angulatus missible oil Celangulin A concentration assay results are as follows:

Celangulin A quality percentage composition in the 1% Celastrus angulatus missible oil

0.35%

Celastrus angulatus cubage in (3) the 1% Celastrus angulatus missible oil

Celastrus angulatus quality percentage composition in the 1% Celastrus angulatus missible oil

1.50%

Claims

1. Celastrus angulatus mother liquor contains the active component of following quality percentage composition: Celangulin A1.9 ~ 3.5%, Celangulin B3.0 ~ 5.0%, Celangulin G2.5 ~ 4.0%, Celangulin E2.50 ~ 4.10%; Described active component is 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound.

2. according to claim 1 Celastrus angulatus mother liquor, it is characterized in that, the active component that wherein also contains following quality percentage composition: Celangulin C, Celastrus angulatus II and Celangulin F1.2 ~ 3.5%, Celastrus angulatus IV0.3 ~ 1.0%, Celangulin A, Celangulin E and Celastrus angulatus C3.0 ~ 4.5%, Celangulin H0.3 ~ 2.0%; Preferably, wherein Celangulin C is 0.35 ~ 1.40%, and Celangulin F is 0.10 ~ 0.90%, and Celastrus angulatus II is 0.60 ~ 1.21%, and Celastrus angulatus C is 0.30 ~ 0.55%.

3. Celastrus angulatus mother liquor according to claim 1 and 2 is characterized in that, the total quality percentage composition of 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound is not less than 10%; Be preferably 11.5 ~ 18.5%.

4. the preparation method of the described Celastrus angulatus mother liquor of claim 1 ~ 3 any one, comprise: take by weighing the Celastrus angulatus root or/and skin, pulverize, the benzene that with volume is 5 ~ 7 times extracts 1 ~ 2 time in 45 ~ 55 ℃ of lower ultrasonic circulations, the each extraction 35 ~ 55 minutes, reduced pressure concentration became medicinal extract after filtrate merged, and namely got the Celastrus angulatus mother liquor.

5. a Celastrus angulatus aqueous emulsion comprises the described Celastrus angulatus mother liquor of claim 1 ~ 3 any one.

6. Celastrus angulatus aqueous emulsion according to claim 5 is characterized in that, comprises the composition of following weight portion: 15 ~ 20 parts in described Celastrus angulatus mother liquor, 8 ~ 12 parts of emulsifier, 3 ~ 4 parts of antifreezes.

7. Celastrus angulatus aqueous emulsion according to claim 6 is characterized in that, described emulsifier is: alkylaryl polyoxyethylene poly-oxygen propylene aether or alkylaryl polyoxyethylene APEO; Described antifreeze is: ethylene glycol or propane diols.

8. the described Celastrus angulatus aqueous emulsion of any one is characterized in that according to claim 5 ~ 7, and total quality percentage composition of 4-OH-beta-dihydroagarofuran sesquiterpene polyolester compound is not less than 2% in this Celastrus angulatus aqueous emulsion; Be preferably 2.3 ~ 3.0%.

9. the quality determining method of a Celastrus angulatus mother liquor may further comprise the steps:

2) Celastrus angulatus mother liquor feature extraction thing is carried out IGD carbon-13 nmr spectra finger-print and detect, obtain several active component characteristic peak peak intensities in the described Celastrus angulatus mother liquor feature extraction thing according to finger-print; And determine the characteristic peak peak intensity of the corresponding criterion referenced product of described each active component with same way as;

4) utilize ratio and the described absolute content of the characteristic peak peak intensity of each active component characteristic peak peak intensity and corresponding criterion referenced product, calculate the content of each active component in the Celastrus angulatus mother liquor and the content of active component group.

10. quality determining method according to claim 9, it is characterized in that, the preparation method of described Celastrus angulatus mother liquor feature extraction thing, comprise: take by weighing the Celastrus angulatus mother liquor, filter rear at 55 ~ 65 ℃ of lower reduced pressure concentrations, reclaim solvent to the medicinal extract shape, medicinal extract with 6 ~ 10 times of amount volumes, the dissolving of 80% ~ 90% methyl alcohol after, use again the petroleum ether extraction 1 ~ 2 time of 3 ~ 5 times of amount volumes, each extraction 20 ~ 40 minutes, reduced pressure concentration after methanol layer filters reclaims solvent to doing, and namely gets Celastrus angulatus mother liquor feature extraction thing.

11. according to claim 9 or 10 described quality determining methods, it is characterized in that step 2) in, the active component characteristic peak in the described Celastrus angulatus mother liquor feature extraction thing is: C-15 absworption peak, its chemical shift are δ _C60.0 ~ 66.0; Preferably, select C-8 further to distinguish Celangulin A, Celangulin E, Celastrus angulatus C as characteristic peak, the chemical shift of C-8 is δ _C73.3 ~ 74.7; Select C-11 further to distinguish Celangulin C, Celangulin F and Celastrus angulatus II as characteristic peak, the chemical shift of C-11 is δ _C82.5 ~ 83.9.

12. the described quality determining method of any one is characterized in that according to claim 9 ~ 11, in the step 3), the absolute content of described criterion referenced product refers to: use the Celastrus angulatus mother liquor Plays of quantitative analysis means mensuration with reference to the quality percentage composition of product; Described quantitative analysis means are high performance liquid chromatography.

13. the described quality determining method of any one is characterized in that according to claim 9 ~ 12, described criterion referenced product are Celangulin A.

14. the described quality determining method of any one is characterized in that according to claim 9 ~ 13, in the step 4), the coupling formula that calculates the content of each active component is:

X_{n} = \frac{X_{1} M_{n} h_{n}}{M_{1} h_{1}};

Wherein:

X ₁Be the absolute content of step 3) with the criterion referenced product that a certain active component is corresponding in the Celastrus angulatus mother liquor of quantitative analysis means mensuration;

M _nMolecular weight for a certain active component;

15. quality determining method according to claim 14 is characterized in that, calculates the coefficient of each active component in the Celastrus angulatus mother liquor and the formula of overall coefficient to be:

F_{n} = \frac{M_{n} h_{n}}{M_{1} h_{1}};

F wherein _nRatio coefficient for the criterion referenced product quality percentage composition that a certain active component is corresponding with it in the Celastrus angulatus mother liquor; M ₁, h ₁, M _nAnd h _nImplication with the corresponding definition in the claim 14, overall coefficient refers to the coefficient F of each active component _nSum.

16. the quality determining method of a Celastrus angulatus aqueous emulsion, utilize the described method of claim 9 ~ 15 any one to detect the content that obtains each active component in the Celastrus angulatus mother liquor, recycle the quality percentage composition that following formula to calculating goes out each active component in the Celastrus angulatus aqueous emulsion: W _n=W ₁F _nWherein:

W ₁For measure the absolute content of the criterion referenced product that a certain active component is corresponding in the Celastrus angulatus aqueous emulsion that obtains with the quantitative analysis means;

F _nRatio coefficient for the criterion referenced product quality percentage composition that a certain active component is corresponding with it in the described Celastrus angulatus mother liquor of claim 15;

The content of active component group is each active component W in the Celastrus angulatus aqueous emulsion _nSum; This Celastrus angulatus aqueous emulsion is prepared by the Celastrus angulatus mother liquor.