CN103760185A - Method for identifying neem oil - Google Patents

Method for identifying neem oil Download PDF

Info

Publication number
CN103760185A
CN103760185A CN201410039926.1A CN201410039926A CN103760185A CN 103760185 A CN103760185 A CN 103760185A CN 201410039926 A CN201410039926 A CN 201410039926A CN 103760185 A CN103760185 A CN 103760185A
Authority
CN
China
Prior art keywords
nim oil
active component
peak
product
feature extraction
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.)
Granted
Application number
CN201410039926.1A
Other languages
Chinese (zh)
Other versions
CN103760185B (en
Inventor
赵天增
张海艳
董建军
景炳年
陈玲
常霞
范毅
张义东
魏悦
于立芹
刘雨晴
李自红
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.)
Henan Kegao Vegetable Natural Product Development Engineering Technology Co ltd
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 CN201410039926.1A priority Critical patent/CN103760185B/en
Publication of CN103760185A publication Critical patent/CN103760185A/en
Application granted granted Critical
Publication of CN103760185B publication Critical patent/CN103760185B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Landscapes

  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)

Abstract

The invention relates to a method for identifying neem oil. The method comprises the following steps: (1) the neem oil or neem oil derivative is directly taken as a feature extract containing a fatty acid active component group; (2) IGD nuclear magnetic resonance carbon spectrum fingerprint detection is implemented for the feature extract, and the peak strengths of a plurality of active component feature peaks in the feature extract are obtained according to the fingerprint; the peak strengths of feature peaks of standard reference substances corresponding to the active components are measured in the same manner; (3) the absolute contents of the standard reference substances are obtained through a quantitative analysis method; (4) the contents of all active components in the neem oil or neem oil derivative and the content of the active component group are calculated by using the ratio value of the peak strengths of the feature peaks of all active components to the peak strengths of the feature peaks of corresponding standard reference substances and the absolute contents of the standard reference substances. The method can measure what fatty acid compounds are contained in the neem oil and the content ratios of the fatty acid compounds so as to achieve the purpose of identifying the quality of the neem oil.

Description

A kind of method of differentiating nim oil
Technical field
The invention belongs to the discriminating field of natural medicinal plant, particularly, relate to a kind of method of differentiating nim oil.
Background technology
Nim oil is the vegetable oil squeezing from print chinaberry (Azadirachta indica A.Juss) fruit, having food refusal repellent, reduce insect gut vigor, suppress insect growth growth, suppress and stop insect molting, suppress the effects such as adult mate and oviposit, is the desirable feedstock of manufacturing each biological pesticide.Have that insecticidal spectrum is wide, insect is difficult for developing immunity to drugs, the feature such as noresidue in environment, can with Multiple Pesticides and fungicide compounding.Excessive in India and Bangladesh etc., nim oil is the most often applied to the nursing maintenance products such as soap, hair products, body lotion and hand lotion.The scope that nim oil is used is from disease of skin, infection, inflammation disease, fever, is even applied to rheumatoid releive etc.
Nim oil complicated component, each component analysis is more difficult.Conventionally, people carry out to nim oil the total fatty acids methyl esters that saponification has obtained nim oil, then with GC or the wherein relative content of contained higher fatty acid [1. sieve happiness honor of GC-MS qualitative determination, Deng. Chinese grease 2002,27 (4): 78. 2. Li Jun peaks, Deng. Yunnan Institute for nationalities's journal 2005,14 (3): 278.].Wherein main stream approach GC chromatography is owing to being subject to non-chromatographic condition (as chromatographic column internal diameter, length, the fixing phase trade mark, carrier granularity, flow rate of mobile phase, mixed flow each component ratio, column temperature, sample size, detector sensitivity etc. mutually) impact is large and needing by standard items etc. reason, and repeatability and feasibility all exist many limitation; There is the problems such as MS degree of ionization and matrix interference in GC-MS analytical approach.
IGD carbon-13 nmr spectra coupling (IGD 13c NMR coupling) 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 ( 1h NMR) fingerprint pattern technology [Zhao Tianzeng, in .1HNMR fingerprint technique plant identification Chinese medicine, Chinese herbal medicine 2000,31 (11): 868-870] on basis, combine other technologies (such as current most widely used high efficiency liquid phase (HPLC) fingerprint pattern technology [Xie Peishan etc., chromatographic fingerprints of Chinese materia medica, People's Health Publisher, 2005] a kind of new comprehensive fingerprint pattern technology of non-single means) proposing.
Research and the application of nim oil IGD carbon-13 nmr spectra coupling finger-print, to the discriminating of nim oil and evaluation, provide a new thinking, also for strengthening systematization and the standardization of the inherent composition Study of nim oil, realize the assurance that science is provided in line with international standards.Along with this technology applying in other Chinese crude drugs and extract thereof, botanical pesticide, the great scientific value of this technology will be increasingly outstanding.
Summary of the invention
For solving the problem of prior art, the object of the invention is to provide a kind of method of differentiating nim oil.
To achieve these goals, the present invention adopts following technical scheme:
A method of differentiating nim oil, it comprises the following steps:
1) get nim oil or nim oil spin-off directly as the feature extraction thing that contains fatty acid active component group;
2) described feature extraction thing is carried out to IGD carbon-13 nmr spectra finger-print and detect, according to finger-print, obtain several active component characteristic peak peak intensities in feature extraction thing; And determine described each active component respective standard with reference to the characteristic peak peak intensity of product by same way (IGD carbon-13 nmr spectra finger-print);
3) by quantitative test means, obtain described standard with reference to the absolute content of product;
4) utilize each active component characteristic peak peak intensity and respective standard with reference to the ratio of the characteristic peak peak intensity of product and described standard the absolute content with reference to product, calculate the content of each active component and the total content of this active component, the i.e. content of active component group in nim oil or nim oil spin-off.
Wherein, step 2) in, feature extraction thing being carried out to IGD carbon-13 nmr spectra finger-print and detect, the solvent of dissolving characteristic extract is deuterochloroform (CDCl 3); The mass volume ratio of feature extraction thing and described solvent is 190:1-210:1(mg:ml), preferred 200:1.
Step 2) in, the active component characteristic peak in feature extraction thing is carbonyl absorption peak (being C-1 absorption peak), and its chemical shift is δ c170.0-185.0.
Step 2), in, described peak intensity can adopt peak height method, area integral method or gravimetric method to calculate.
In step 3), described quantitative test means are gas chromatography (GC) method; Described standard refers to reference to the absolute content of product: the standard of measuring by quantitative test means is with reference to the quality percentage composition of product.
Further, the standard of the active component in described feature extraction thing is methyl oleate with reference to product.
Wherein, in step 4), the coupling formula that calculates each active component content is:
; Wherein:
W 1the standard that in the nim oil of measuring by quantitative test means for step 3) or nim oil spin-off, a certain active component is corresponding is with reference to the absolute content of product;
M 1for standard that in described nim oil or nim oil spin-off, a certain active component is corresponding is with reference to carbon number corresponding to peak of the molecular weight of product/quantitatively;
H 1for standard that in the nim oil by IGD carbon-13 nmr spectra determining fingerprint pattern or nim oil spin-off, a certain active component the is corresponding characteristic peak peak intensity with reference to product;
W nquality percentage composition for a certain active component in nim oil or nim oil spin-off;
M ncarbon number corresponding to molecular weight/quantitative peak for a certain active component in nim oil or nim oil spin-off;
H ncharacteristic peak peak intensity for a certain active component in the nim oil by IGD carbon-13 nmr spectra determining fingerprint pattern or nim oil spin-off; 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:
Figure 247097DEST_PATH_IMAGE002
Figure 2014100399261100002DEST_PATH_IMAGE003
Figure 101921DEST_PATH_IMAGE004
Figure 2014100399261100002DEST_PATH_IMAGE005
Active component group described in the inventive method, refers to the summation of fatty acid active component.
In the present invention, four steps that nim oil is differentiated are an integral body, indispensable; Four steps have distinctive feature separately; If separately or simple combination can not detect ratio and the content of complicated ingredient in medicinal plant.Nim oil contains multiple fatty acid or ester constituents, and its analyzing and testing is a difficult problem always.The qualitative and quantitative analysis aspect that currently available technology solves differential plant kind and evaluation plant source product quality exists significant limitation.Fact proved, IGD carbon-13 nmr spectra coupling fingerprint pattern technology solves the key of existing issue just.
IGD 13c NMR coupling finger-print is the mixed spectrum of a plurality of active components, unavoidably can cause the crowded, even overlapping of excellent peak one by one.In order to make result of calculation accurate, it is necessary selecting each specific characteristic peak, active component carbon peak in active component group that chemical shift difference is larger.Dissimilar compound (fatty acid or ester constituents) carbon spectral difference is very not large, and the selection of characteristic peak can be ever-changing; Same type compound carbon spectral difference is not very little, and a lot of peak overlaps are serious, need have deep nuclear magnetic resonance spectrum knowledge, through in many ways comparing, turning over, could select good characteristic peak.And characteristic peak selection is bad, have no idea active component group to carry out accurate quantitative analysis analysis this problem that also the application need to solve just.According to the feature of fatty acid ester composition in nim oil, what characteristic peak was selected is C-1 carbonyl absorption peak.So, according to the feature of different activities composition, need to select different specific characteristic peaks, active component carbon peak, this is the application's one of innovative point just.
Because characteristic peak chemical shift difference is very little, in a lot of situations, only after radix point the 1st show difference, so the sequence of characteristic peak is the key of determining main active and ratio thereof, there is no deep nuclear magnetic resonance spectrum knowledge and separated basis, be difficult to determine active component and the ratio thereof of characteristic peak representative, also just cannot carry out accurate qualitative and quantitative analysis to active component group, according to the feature of fatty acid ester composition in nim oil, by characteristic peak, determine concrete fatty acid or ester constituents, also acid or ester constituents can be distinguished; This is also one of innovative point of the application.
It is the selection of choice criteria product and quantitative analysis tech that key is calculated in coupling.Analyze quantitative means and can select high efficiency 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, can be also additional with reference to product as external standard.According to the feature of composition in nim oil, what analyze quantitative means selection is gas chromatography, what standard was selected with reference to product is that methyl oleate is (because oleic acid boiling point is too high, gas phase is generally by after methyl oleate, measure the wherein content of methyl oleate, but in nim oil, oleic acid physical presence form is triglyceride oleate).This is one of difficult point of the application, is also one of innovative point of the application simultaneously.
The content of each active component of the present invention and the total content of this active component calculate, and are 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 13c NMR coupling finger-print has following feature:
1. stability (repeatability): IGD 13the chemical shift data that C NMR obtains is second after radix point, and explanation property is good, reproducible; The non-chromatographic condition of GC (as chromatographic column internal diameter, length, the fixing phase trade mark, carrier granularity, flow rate of mobile phase, mixed flow each component ratio, column temperature, sample size, detector sensitivity etc. mutually) changes etc., the retention time data variation obtaining 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 sample in C NMR finger-print; There is not this relation in HPLC, GC, UV, IR, MS.
3. reliability (unicity): IGD 13c NMR spectrum peak and carbon on different activities composition in sample and different group thereof are strict one-to-one relationships; There is 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, can belong to each the carbon peak in collection of illustrative plates; HPLC, GC need a plurality of reference substances; IR is difficult for resolving; UV quantity of information is few; MS has the problems such as degree of ionization and matrix interference.
IGD carbon-13 nmr spectra finger-print can only show in feature extraction thing, there is which active component, and the quantitative ratio between these active components, and the absolute content of these active components must be analyzed quantitative means with reference to product and other by standard, then obtain by coupling formula.The present invention adopts IGD carbon-13 nmr spectra finger-print to differentiate nim oil, can reflect in nim oil and contain which fatty acid compound and the ratio between them, reaches the object to nim oil Quality Identification.The range of linearity is wide, highly sensitive, and repeatability and feasibility are good.
Accompanying drawing explanation
Fig. 1-a is the IGD carbon-13 nmr spectra finger-print of nim oil products A in embodiment 1;
Fig. 1-b is that the part, IGD carbon-13 nmr spectra Fingerprints peak of nim oil products A in embodiment 1 widens enlarged drawing;
Fig. 2-a is the IGD carbon-13 nmr spectra finger-print of nim oil product B in embodiment 2;
Fig. 2-b is that the part, IGD carbon-13 nmr spectra Fingerprints peak of nim oil product B in embodiment 2 widens enlarged drawing;
Fig. 3-a is the IGD carbon-13 nmr spectra finger-print of nim oil products C in embodiment 3;
Fig. 3-b is that the part, IGD carbon-13 nmr spectra Fingerprints peak of nim oil products C in embodiment 3 widens enlarged drawing.
Embodiment
By specific embodiment, technical scheme of the present invention is described below, but protection scope of the present invention is not limited to this.
one, nim oil IGD carbon-13 nmr spectra finger-print research
(1) acquisition of fatty acid feature extraction thing (CET)
Directly get nim oil as the feature extraction thing that contains fatty acid active component group.
(2) the IGD carbon-13 nmr spectra finger-print of fatty acid feature extraction thing detects
Get fatty acid feature extraction thing 100 mg, be dissolved in 0.5mL CDCl 3in, make IGD carbon-13 nmr spectra finger-print and detect, obtain fatty acid IGD carbon-13 nmr spectra finger-print.
(3) the IGD carbon-13 nmr spectra finger-print of fatty acid feature extraction thing is resolved
1. differentiate
In fatty acid feature extraction thing IGD carbon-13 nmr spectra finger-print, clearly illustrate the characteristic signal of fatty acid compound.
2. each active component characteristic peak in fatty acid feature extraction thing is chosen
Owing to containing a series of fatty acid and ester type compound thereof in fatty acid feature extraction thing, carbon peak intersects morely.In order to measure the ratio of each active component, must select respective peaks that chemical shift difference is larger as characteristic peak., through reality, investigate for this reason, select δ cone group of C-1 carbonyl carbon peak of 170.0-185.0 left and right is as this specific characteristic peak, active component carbon peak, and it is former because C-1 is carbonyl carbon, larger, easy to identify with other carbon geochemistry displacement difference; Between carbonyl carbon peak, different Compound C-1, chemical shift also has certain difference.
3. standard is with reference to the selection of product
Triglyceride oleate is one of main active of liposoluble constituent in fatty acid feature extraction thing, and its carbonyl carbon chemical shift is about δ c173.2 (C-1''), 173.1 (C-1'), 172.8 (C-1), δ c172.8 (C-1) do not have overlapping with other main fatty acid composition at this.Triglyceride oleate saponification process after the overwhelming majority become methyl oleate because select methyl oleate as standard with reference to product.
(4) adopt GC method to measure the content of oleic acid in nim oil
1) chromatographic condition
With TC-FFAP quartz capillary column (thickness 0.4 μ M, 600 * 0.25mm) post, 60～220 ℃ of column temperature scopes, 3 ℃/min of temperature programmes, 220 ℃ of stable 30min, injection port and detector temperature are 220 ℃.Use helium as carrier gas, flame ionization ditector detects.Test, through the nim oil of the direct esterification of 0.4 mol/l KOH methanol solution, is usingd methyl oleate as reference.
2) triglyceride oleate absolute content calculates
1. by following formula, calculate methyl oleate mass concentration in need testing solution:
Figure 65679DEST_PATH_IMAGE006
C x: methyl oleate mass concentration (ug/mL) in nim oil need testing solution;
C r: standard is with reference to product solution methyl oleate mass concentration (ug/mL);
A x: methyl oleate peak area in the nim oil need testing solution of being measured by GC;
A r: the standard of being measured by GC is with reference to product solution methyl oleate peak area.
2. by following formula, calculate methyl oleate quality percentage composition in nim oil:
Figure 2014100399261100002DEST_PATH_IMAGE007
W' methyl oleate(%): methyl oleate quality percentage composition in nim oil;
C x: methyl oleate mass concentration (ug/mL) in nim oil need testing solution;
M test sample: the nim oil need testing solution quality (mg) taking.
3. triglyceride oleate quality percentage composition calculates
The existence form of oleic acid in nim oil is triglyceride oleate, and adding existence form after the esterification of KOH methanol solution is methyl oleate.Therefore the content that, records methyl oleate in the need testing solution after esterification is triglyceride oleate quality percentage composition in nim oil.
(5) by coupling formula, calculate fatty acid each main active content and total amount, the i.e. content of fatty acid active component group in nim oil
Figure 398571DEST_PATH_IMAGE008
W 1: the nim oil Plays that GC method is measured is with reference to the quality percentage composition of product triglyceride oleate;
M 1: standard is with reference to the carbon number corresponding to molecular weight/quantitative peak of product triglyceride oleate;
H 1: standard is with reference to the characteristic peak peak intensity (peak height) of product triglyceride oleate;
W n: the quality percentage composition of a certain fatty acid active component in the nim oil that GC method is measured;
M n: carbon number corresponding to the molecular weight of a certain fatty acid active component in nim oil/quantitative peak;
H n: the characteristic peak peak intensity (peak height) of a certain fatty acid active component in nim oil.
two, instrument, reagent and material
Key instrument and equipment
Nuclear magnetic resonance spectrometer Bruker DPX 400 types.
Gas chromatograph: Shimadzu GC-2010 (FID hydrogen flame ionization detector) type.
Nim oil products A (Yunnan Nan Bao biotechnology Ltd), nim oil product B (bright Neem industry is developed incorporated company), nim oil products C (Xi'an Tian Yi Bioisystech Co., Ltd); Methyl oleate, chemical reference substance (Sen Beijia bio tech ltd, Nanjing); Reagent: methyl alcohol (chromatographically pure, Tianjin Siyou Fine Chemicals Co., Ltd.), methyl alcohol (analyze pure, Tianjin Chemical Reagents Factory No.1).
three, fatty acid carbons spectrum data
Triglyceride oleate (Glycerol trioleate) C 57h 104o 6
13CNMR(CDCl 3,100MHz)δ C:173.28(C-1', 1''), 172.83(C-1), 130.20, 129.67, 68.85, 62.07, 34.17, 34.03, 32.40, 31.90, 31.51, 30.41, 29.68, 29.32, 29.50, 29.31, 29.10，27.17, 25.60, 24.84, 22.67, 14.09
Figure 2014100399261100002DEST_PATH_IMAGE009
。
Monoolein (Glycerol oleate) C 21h 40o 4
13CNMR(75MHz,CDCl 3)δ C:173.14(C-1),33.88(C-2),24.42(C-3),29.10(C-4),29.61(C-5),29.54(C-6),29.30(C-7),27.21(C-8),130.51(C-9),128.89(C-10),24.77(C-11),29.31(C-12),29.55(C-13),26.77(C-14),29.31(C-15),31.87(C-16),22.66(C-17),14.08(C-18),68.83(C-a),62.04(C-b,C-c)
。
Linoleic acid glyceryl ester (Glycerol trilinolet) C 57h 98o 6
13CNMR(150MHz,CDCl 3)δ C:173.16(C-1''),173.05(C-1'),172.75(C-1), 34.11(C-2'')，34.92（C-2'）, 33.86(C-2), 24.81(C-3''), 24.41(C-3', C-3), 29.29(C-4, C-4', C-4''), 29.61(C-5, C-5', C-5''), 29.53(C-6, C-6', C-6''), 29.64(C-7, C-7', C-7''), 27.20(C-8'', C-8'), 27.13(C-8), 130.01(C-9''), 129.89(C-9', C-9), 128.01(C-10''), 127.82(C-10', C-10), 25.56(C-11''), 24.81(C-11', C-11), 128.87(C-12, C-12', C-12''), 130.46(C-13, C-13', C-13''), 26.74(C-14, C-14', C-14''), 29.53(C-15, C-15', C-15''), 31.87(C-16, C-16''), 31.47(C-16), 22.64(C-17'', C-17'), 22.52(C-17), 14.05(C-18'', C-18'), 14.01(C-18), 68.82(C-a),62.04(C-b,C-c)
Figure 2014100399261100002DEST_PATH_IMAGE011
。
Monopalmitin (Glyceryl monopalmitate) C 19h 38o 4
13CNMR(100MHz,CDCl 3)δ C: 65.2(C-a),70.4(C-b),63.4(C-c),174.4(C-1), 34.2(C-2), 31.9(C-3), 29.7-29.1 (C-4 to C-13), 24.9(C-14), 22.7(C-15), 14.0(C-16).
Figure 160040DEST_PATH_IMAGE012
。
Tristerin (glycerol-1-ocatadecanoate) C 21h 42o 4
13CNMR(CDCl 3, )δ C:65.10(C-a), 70.29(C-b), 63.50(C-c), 174.48(C-1), 32.03(C-2), 22.78(C-3), 29.46(C-4), 29.81(C-5～C-15), 22.78(C-17), 14.18(C-18)
。
2-stearic acid-4-hexadecylic acid-2,4-diamyl ester (2-Octadecanoic acid-4-palmitic acid-2,4-pentanediyl ester) C 39h 76o 4
13CNMR(CDCl 3,50MHz)δ C: 14.03(C-a), 68.33(C-b), 31.84(C-c), 68.33(C-d), 14.03 (C-e), 173.83(C-1,1'), 64.95(C-2,2'), 34.02(C-3,3'), 24.81(C-4,4'), 29.04-29.61 (C-5,5'), 22.61(C-6,6'), 14.03(C-7,7')
Figure 2014100399261100002DEST_PATH_IMAGE015
。
18 carbonic acid-tetrahydrofuran-3,4-diester (Octadecanoic acid-tetrahydrofuran-3,4-diyl ester) C 40h 76o 5636
13CNMR(CDCl 3,50MHz) δ C: 65.05(C-b, e), 68.45(C-c, d), 173.89 (C-1, 1'), 34.11 (C-2, 2'), 24.90 (C-3,3'), 29.13 (C-4,4'), 31.92 (C-5,5'), 22.68 (C-6, 6'), 14.09(C-7,7')
Figure DEST_PATH_IMAGE017
。
Ethyl palmitate (ethyl palmitate) C 18h 36o 2
13CNMR(CDCl 3,100MHz)δ C: 173.33(C-1), 29.71(C-2～C-12), 29.38(C-13), 31.94(C-14), 22.17(C-15), 14.13(C-16), 66.10(OCH 2)
Figure DEST_PATH_IMAGE019
。
Methyl hexadecanoate (methyl palmitate) C 17h 34o 2
13CNMR(CDCl 3,125MHz)δ C：55.5(OCH 3), 174.1(C-1), 31.9(C-2), 29.4(C-3to14), 22.7(C-15), 14.1(C-16)
Figure DEST_PATH_IMAGE021
Methyl arachidate (Methyl Arachidate) C 21h 42o 2
13CNMR(CDCl 3,100MHz)δ C：174.5(C-1),51.5(OCH 3),34.0(C-2), 24.9(C-3), 29.0-29.7(C-4-C-17), 31.9(C-18), 22.7(C-19),14.1(C-20)
Figure DEST_PATH_IMAGE023
。
Lignoceric acid methyl esters (Lianoceric acid methyl ester) C 25h 50o 2
13CNMR(CDCl 3,100MHz)δ C：174.3（C-1）, 51.4(OCH 3),34.1(C-2)，31.9（C-3）, 29.4(C4-C21), 24.9(C-22), 22.7(C-23), 14.1(C-24)
Figure DEST_PATH_IMAGE025
。
Heptadecanoic acide (heptadecanoic acid) C 17h 34o 2
13CNMR(CDCl 3,150MHz)δ C:179.92(C-1),34.02(C-2),24.67(C-3),29.68,29.68,29.66,29.65,29.63,29.58,29.42,29.35,29.23,29.05(C-4-C-14),31.92(C-15), 22.68(C-16),14.10(C-17)
Figure DEST_PATH_IMAGE027
。
Stearic acid (octadecanoid acid, Stearic acid) C 18h 36o 2
13CNMR(CDCl 3,100MHz)δ C:180.1(C-1),34.2(C-2),24.7(C-3),29.2(C-4),29.7(C-6～C-14),31.9(C-16),22.7(C-17),14.4(C-18)
Figure DEST_PATH_IMAGE029
。
Palmitic acid (palmitic acid, palmitic acid) C 16h 32o 2
13CNMR(CDCl 3,100MHz)δ C: 179.5(C-1), 29.0-29.5 (C-2～C-13) 12, 31.9 (C-14) ,22.7 (C-15), 14.1(C-16)
。
embodiment 1: the IGD carbon-13 nmr spectra coupling finger-print of nim oil products A
(1) feature extraction thing preparation
Select nim oil products A directly as fatty acid feature extraction thing.
(2) feature extraction thing IGD carbon-13 nmr spectra finger-print detects
Get fatty acid feature extraction thing 100mg, be dissolved in 0.5mL CDCl 3in, make IGD carbon-13 nmr spectra and detect, both obtained the sour feature extraction thing IGD carbon-13 nmr spectra finger-print of nim oil fat.
(3) the sour feature extraction thing IGD carbon-13 nmr spectra finger-print of nim oil fat
1) IGD carbon-13 nmr spectra finger-print is differentiated
In the IGD carbon-13 nmr spectra finger-print of the fatty acid feature extraction thing of nim oil products A, clearly illustrate the characteristic signal of fatty acid compound.Fatty acid compound triglyceride oleate, linoleic acid glyceryl ester, 18 carbonic acid-tetrahydrofuran-3,4-diester, 1.2-stearic acid-4-hexadecylic acid-2,4-diamyl ester, palmitic acid, stearic acid 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 part widens enlarged drawing and sees accompanying drawing 1-b.
2) in fatty acid feature extraction thing, each active component ratio measuring result is as follows:
Figure DEST_PATH_IMAGE033
3) nim oil product ait is as follows that middle triglyceride oleate quality percentage composition is measured (GC method) result:
Figure DEST_PATH_IMAGE035
4) nim oil product amiddle fatty acid active component quality percentage composition measurement result is as follows:
embodiment 2: IGD carbon-13 nmr spectra-GC coupling finger-print of nim oil product B
(1) feature extraction thing preparation
Select nim oil product B directly as fatty acid feature extraction thing.
(2) feature extraction thing IGD carbon-13 nmr spectra finger-print detects
Get fatty acid feature extraction thing 100mg, be dissolved in 0.5mL CDCl 3in, make IGD carbon-13 nmr spectra and detect, obtain fatty acid feature extraction thing IGD carbon-13 nmr spectra finger-print.
(3) fatty acid feature extraction thing 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 the fatty acid feature extraction thing of nim oil product B, clearly illustrate the characteristic signal of fatty acid compound.Fatty acid compound triglyceride oleate, linoleic acid glyceryl ester, 18 carbonic acid-tetrahydrofuran-3,4-diester, 1.2-stearic acid-4-hexadecylic acid-2,4-diamyl ester, palmitic acid, stearic acid 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 part widens enlarged drawing and sees accompanying drawing 2-b.
2) in fatty acid feature extraction thing, each active component ratio measuring result is as follows:
Figure DEST_PATH_IMAGE039
3) in nim oil product B, triglyceride oleate quality percentage composition mensuration (GC method) result is as follows:
Figure DEST_PATH_IMAGE041
4) in nim oil product B, fatty acid active component quality percentage composition measurement result is as follows:
Figure DEST_PATH_IMAGE043
embodiment 3: IGD carbon-13 nmr spectra-GC coupling finger-print of nim oil products C
(1) feature extraction thing preparation
Select nim oil products C directly as fatty acid feature extraction thing.
(2) feature extraction thing IGD carbon-13 nmr spectra finger-print detects
Get fatty acid feature extraction thing 100mg, be dissolved in 0.5mL CDCl 3in, make IGD carbon-13 nmr spectra and detect, obtain fatty acid feature extraction thing IGD carbon-13 nmr spectra finger-print.
(3) fatty acid feature extraction thing 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 the fatty acid feature extraction thing of nim oil products C, clearly illustrate the characteristic signal of fatty acid compound.Fatty acid compound triglyceride oleate, linoleic acid glyceryl ester, 18 carbonic acid-tetrahydrofuran-3,4-diester, 1.2-stearic acid-4-hexadecylic acid-2,4-diamyl ester, palmitic acid, stearic acid etc. all have corresponding NMR signal 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 part widens enlarged drawing and sees accompanying drawing 3-b.
2) in fatty acid feature extraction thing, each active component ratio measuring result is as follows:
3) in nim oil products C, triglyceride oleate quality percentage composition mensuration (GC method) result is as follows:
Figure DEST_PATH_IMAGE047
4) in nim oil products C, fatty acid active component quality percentage composition measurement result is as follows:
Figure DEST_PATH_IMAGE049

Claims (6)

1. a method of differentiating nim oil, is characterized in that, comprises the following steps:
1) get nim oil or nim oil spin-off directly as the feature extraction thing that contains fatty acid active component group;
2) described feature extraction thing is carried out to IGD carbon-13 nmr spectra finger-print and detect, according to finger-print, obtain several active component characteristic peak peak intensities in feature extraction thing; And determine described each active component respective standard with reference to the characteristic peak peak intensity of product by same way;
3) by quantitative test means, obtain described standard with reference to the absolute content of product;
4) utilize each active component characteristic peak peak intensity and respective standard with reference to the ratio of the characteristic peak peak intensity of product and described standard with reference to the absolute content of product, calculate the content of each active component and the content of active component group in nim oil or nim oil spin-off.
2. differentiate according to claim 1 the method for nim oil, it is characterized in that step 2) in, feature extraction thing being carried out to IGD carbon-13 nmr spectra finger-print and detect, the solvent of dissolving characteristic extract is deuterochloroform; The mass volume ratio of feature extraction thing and described solvent is 190:1-210:1(mg:ml).
3. according to the method for differentiating nim oil described in claim 1 or 2, it is characterized in that step 2) in, the active component characteristic peak in feature extraction thing is carbonyl absorption peak, its chemical shift is δ c170.0-185.0.
4. differentiate according to claim 1 the method for nim oil, it is characterized in that, in step 3), described quantitative test means are vapor-phase chromatography; Described standard refers to reference to the absolute content of product: the standard of measuring by quantitative test means is with reference to the quality percentage composition of product.
5. differentiate according to claim 1 the method for nim oil, it is characterized in that, the standard of the active component in feature extraction thing is methyl oleate with reference to product.
6. according to the method for differentiating nim oil described in claim 1 to 5 any one, it is characterized in that, in step 4), the coupling formula that calculates each active component content is:
; Wherein:
W 1the standard that in the nim oil of measuring by quantitative test means for step 3) or nim oil spin-off, a certain active component is corresponding is with reference to the absolute content of product;
M 1for standard that in described nim oil or nim oil spin-off, a certain active component is corresponding is with reference to carbon number corresponding to peak of the molecular weight of product/quantitatively;
H 1for standard that in the nim oil by IGD carbon-13 nmr spectra determining fingerprint pattern or nim oil spin-off, a certain active component the is corresponding characteristic peak peak intensity with reference to product;
W nquality percentage composition for a certain active component in nim oil or nim oil spin-off;
M ncarbon number corresponding to molecular weight/quantitative peak for a certain active component in nim oil or nim oil spin-off;
H ncharacteristic peak peak intensity for a certain active component in the nim oil by IGD carbon-13 nmr spectra determining fingerprint pattern or nim oil spin-off.
CN201410039926.1A 2013-01-30 2014-01-27 A kind of method differentiating nim oil Active CN103760185B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201410039926.1A CN103760185B (en) 2013-01-30 2014-01-27 A kind of method differentiating nim oil

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN2013100357245 2013-01-30
CN201310035724.5 2013-01-30
CN201310035724 2013-01-30
CN201410039926.1A CN103760185B (en) 2013-01-30 2014-01-27 A kind of method differentiating nim oil

Publications (2)

Publication Number Publication Date
CN103760185A true CN103760185A (en) 2014-04-30
CN103760185B CN103760185B (en) 2016-03-30

Family

ID=50527460

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410039926.1A Active CN103760185B (en) 2013-01-30 2014-01-27 A kind of method differentiating nim oil

Country Status (1)

Country Link
CN (1) CN103760185B (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040185506A1 (en) * 2003-03-21 2004-09-23 Heavner George A. Epitope mapping using nuclear magnetic resonance
CN101602753A (en) * 2008-06-13 2009-12-16 四川农业大学 Ester compound and its production and application in the nim oil
CN102749348A (en) * 2012-06-18 2012-10-24 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

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040185506A1 (en) * 2003-03-21 2004-09-23 Heavner George A. Epitope mapping using nuclear magnetic resonance
CN101602753A (en) * 2008-06-13 2009-12-16 四川农业大学 Ester compound and its production and application in the nim oil
CN102749348A (en) * 2012-06-18 2012-10-24 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

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
VIRENDRA K DUA ET.AL.: "Larvicidal activity of neem oil (Azadirachta indica) formulation against mosquitoes", 《MALARIA JOURNAL》 *
杜永华: "印楝油体外杀螨活性及其活性物质的分离纯化与结构鉴定研究", 《中国优秀硕士学位论文全文数据库》 *
罗喜荣 等: "印楝籽油脂肪酸组成的分析", 《中国油脂》 *
谭卫红 等: "印楝的化学成分及其生物活性", 《东北林业大学学报》 *

Also Published As

Publication number Publication date
CN103760185B (en) 2016-03-30

Similar Documents

Publication Publication Date Title
Agnolet et al. Comprehensive analysis of commercial willow bark extracts by new technology platform: Combined use of metabolomics, high-performance liquid chromatography–solid-phase extraction–nuclear magnetic resonance spectroscopy and high-resolution radical scavenging assay
CN103913528B (en) Quantitative detection method for pyrethriods pesticides in fresh tea
CN108061768A (en) A kind of structure and its detection method of Radix Paeoniae Alba HPLC characteristic spectrums
Fan et al. Metabolic discrimination of rhizoma Coptidis from different species using 1H NMR spectroscopy and principal component analysis
CN104777249B (en) The method measuring effective ingredient amygdaloside content in cough syrup of loquat leaf
CN104020235B (en) A kind of method of simultaneously measuring Chlorogenic Acid of Flos Lonicerae and galuteolin content
CN106124639A (en) The multicomponent content assaying method of Eucommia ulmoides
CN106353417A (en) UPLC (ultra-performance liquid chromatography) detection method of multiple steroid saponins in Yunnan rhizoma paridis or its polygerm strain
CN103674996B (en) A kind of method differentiating red rooted salvia or spin-off
CN105067747B (en) The detection method of Styrax flavour of a drug ingredients fingerprint and its application in a kind of Heart pill of Musk
CN105548385A (en) Determination method for liquid-phase fingerprint of Xingnaojing injection and standard fingerprint of Xingnaojing injection
CN109991327A (en) One surveys the methods for commenting method evaluation field thistle quality more
CN104374854B (en) A kind of method of multiple phenolic content in HPLC wavelength handoff technique Simultaneously test Noni juice
CN103808749B (en) A kind of method differentiating capsule of weeping forsythia spin-off
CN103913538B (en) The quantitative detecting method of organophosphorus insecticide in a kind of tea fresh leaves
Zhang et al. Simultaneous determination of four active components in Alisma orientale (Sam.) Juz. by HPLC–DAD using a single reference standard
CN103808751B (en) A kind of method differentiating traditional Chinese medicine honeysuckle or spin-off
Gong et al. Novel strategy for quality consistency evaluation of Chinese medicine “YIQING” tablet that combines the simultaneous quantification and screening of ten bioactive constituents
CN108152399A (en) A kind of structure and its detection method of seed of Oriental arborvitae medicinal material UPLC characteristic spectrums
Lü et al. Quality evaluation of Angelica sinensis by simultaneous determination of ten compounds using LC-PDA
CN103134827B (en) Method for identifying angled bittersweet seed medicinal materials
Hou et al. Characteristic chromatogram: a method of discriminate and quantitative analysis for quality evaluation of Uncaria stem with hooks
CN103134877A (en) Method measuring C6-C3 type phenolic acids compound content in tobacco
CN103868943B (en) A kind of method of differentiating Fructus Ligustri Lucidi or spin-off
CN107422047A (en) The discrimination method of Dulong Paris polyphylla

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