CN1844116A - Extraction separation purification and identification of flavonoid monomers in oriental blueberry melanin - Google Patents
Extraction separation purification and identification of flavonoid monomers in oriental blueberry melanin Download PDFInfo
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Abstract
This is a method for flavonoid monomer's extraction, separation, purification and structural identification, belonging to flavonoid field. In this invention, the original source blueburry leaves turn to be blueburry leaves melanin solution after ethanol extraction. Then, macroreticular resinous adsorbent is used for crude separation of flavonoid in melanin solution, polyamide and HW-40 for flavonoid monomer's pufification, high efficiency liquid chromatography-mass spectrum and NMRR for structural identification. Finally, we obtain 7 kinds of flavonoid monomer, named A B C D E F and G. Exact structural identification help to determine B C D E and F is quercetin, chrysin, versulin, Kacmpferol and digitoflavone respectively, and their relative contents are 37.51%,2.26%,9.57%,1.72% and 15.16%.
Description
Technical field
The method that the monomeric extraction of flavonoid compound in a kind of Fructus Vaccinii Bracteati leaf melanochrome, separation, purifying and structure are identified belongs to the flavonoid compound technical field.
Background technology
Fructus Vaccinii Bracteati is a kind of abundant natural resource, and the melanochrome in its leaf has very strong physiological function.Content of flavonoids is very high in Fructus Vaccinii Bracteati leaf melanochrome extract, thus be necessary to its extract, separation, purifying and evaluation, be beneficial to the development and use of Fructus Vaccinii Bracteati.
Extraction separation goes out compound monomer from natural phant, and carries out the Analysis and Identification of structure, is an importance in the natural product research, and meaning is: (1) finds the effective constituent in the medicinal plant, seeks the foundation of its clinical efficacy; (2) help to illustrate the structure activity relationship of natural drug; (3) produce evidence and direction for the physiological Study of the classification of plant, plant.
The separation purification method of flavonoid compound mainly contains various column chromatographies, HPLC method, centrifugal thin layer chromatography, preparation of lamina chromatography and Paper Chromatography etc.Wherein column chromatography is the method for the most frequently used separating and purifying flavone compounds, mainly contain three classes: (1) polyamide column chromatography method: polymeric amide has centrifugation preferably to flavonoid glycoside, its chromatography capacity is big, be suitable for the separation of preparation property, eluent is used water-methanol always, also useful water-ethanol and methyl alcohol-chloroform; (2) silica gel column chromatography: the range of application of this method is the widest, not only can the separating flavone glycosides, also can separate various Flavone aglycones; (3) dextrane gel column chromatography: it mainly is that generally the size order according to molecular weight washes out cylinder when wash-out by molecular sieve effect separating flavone glycoside.The report of HPLC method separating flavone class material is then more, and what be used for the flavonoid compound separation and purification in HPLC generally has a C
8, C
18, the CN post, the different in kind of CN post and preceding two kinds of column packings, polarity is stronger, and is not good for the component separating effect of low polarity part, C
18Post can be realized separating to plant flavone aglycon and aglucon substantially, but it is slow to go out the peak for the big glycosides part wash-out of polarity, and total elution time increases, and separating effect is not very desirable, and C
8Filler polarity is between between the two, and more near C
18Some, thus more satisfactory to the compound separation of flavonoid glycoside.Then several separation and purification that are used for flavonoid compound are then less.In the separation and purification process of flavonoid compound, need sometimes several method is used alternatingly.
The structure identification research of flavonoid compound mainly concentrates on the aglycon part, and the basic structure of aglycon can be divided into the main type of kind more than 10.The aglycon of flavonoid compound is made up of 15 carbon atoms, and total A, B, a C3 ring are formed, and structure is as follows.Common substituting group has hydroxyl, methoxyl group, isopentene group or the like on A, B ring.
Mainly utilize spectral method for flavonoid compound structural research at present, comprising: mass spectrum, UV spectrum and nuclear magnetic resonance map.When research flavonoid compound aglycon structure, utilize mass spectrum more effective, because the molecular ion peak of flavonoid compound aglycon is very strong, and owing to the glycosyl that is connected fragment appears, so research is difficult a little in the research flavonoid glycoside compound.Flavonoid compound is having two absorption bands between 300～400nm and between 240～280nm, the former is the absorption by the transition of electron generation of B ring atom, and the latter is the absorption by the transition of electron generation of A ring atom.Add different chemical reagent (sodium methylate, sodium ethylate, aluminum chloride, boric acid, sodium-acetate etc.), two absorption bands of this of flavonoid compound can since the variation of aglycon part-structure produce mobile, so can utilize the different mobile reagent of interpolation to judge substituting group and position thereof on the aglycon.Utilize the structure of nuclear magnetic resonance technique research flavonoid compound generally have proton nmr spectra (
1H-NMR), carbon-13 nmr spectra (
13C-NMR) and two dimensional NMR spectrum (2D-NMR) or the like.
Summary of the invention
The purpose of this invention is to provide the method that the monomeric extraction of flavonoid compound in a kind of Fructus Vaccinii Bracteati leaf melanochrome, separation, purifying and structure are identified, utilize method separation and purification from Fructus Vaccinii Bracteati leaf melanochrome such as macroporous adsorbent resin, polyamide column, HW-40 post to obtain the flavonoid compound monomer, utilize HPLC-MS and nuclear magnetic resonance technique to carry out structural research.
Technical scheme of the present invention:
With the Fructus Vaccinii Bracteati leaf is raw material, obtain Fructus Vaccinii Bracteati leaf melanochrome solution through extraction using alcohol, through macroporous adsorbent resin the flavonoid compound in the melanochrome is carried out roughing out again, by polyamide column and HW-40 post the flavonoid compound monomer is carried out purifying again, with high performance liquid chromatography-mass spectrum, nuclear magnetic resonance technique purified components is carried out structure again and identify.
(1) extraction of Fructus Vaccinii Bracteati leaf melanochrome solution: will clean up, air dried Fructus Vaccinii Bracteati leaf utilizes tissue mashing machine's wet method to smash to pieces, add reactor, utilize spirituous solution to extract, solid-liquid ratio is 1: 20～1: 30 in mass, the spirituous solution volumetric concentration is 60%, extracting temperature is 55～60 ℃, stir and extract 1h, solution is leached, obtain the aqueous solution of extract behind the recovered alcohol, the spirituous solution adjusted volume concentration that reclaims is to be added to reactor after 60%, to the leaf second extraction 1h in the reactor, solution is leached the aqueous solution merging that obtains the aqueous solution of extract behind the recovered alcohol and extract the extract that obtains for the first time according to the same terms, the moisture content that is concentrated into extracting solution is no more than 10%, and is standby.
(2) macroporous adsorbent resin carries out roughing out to the flavonoid compound in the melanochrome: add volume ratio under the extracting solution normal temperature and be 1: 1 n-butanol extraction half an hour, butanol extraction liquid is evaporated to does not have the alcohol flavor, the adding by volume is 1: 2～1: 3 a deionized water, sonic oscillation, AB-8 macroporous adsorptive resins on the aqueous solution, first deionized water wash-out with 2～3 times of column volumes, discard elutriant, be 95% alcohol wash-out again with the volumetric concentration of 3～5 times of column volumes, collect elutriant, be concentrated into ethanol content and be no more than 5%, standby.
(3) polyamide column and HW-40 post carry out purifying to the flavonoid compound monomer: the wash-out concentrated solution that macroporous adsorptive resins is eluted, last polymeric amide huge pillar, column volume φ 4cm * 50cm, the difference water, volumetric concentration is 20% alcohol, 45% alcohol, 70% alcohol, 95% alcohol carries out wash-out successively, each elutriant consumption is 2～3 column volumes, collect elutriant, point sample analysis on thin plate, merge identical component, same composition is concentrated into moisture content and is no more than 5%, and each component goes up the polymeric amide pillar respectively, φ 3cm * 30cm, increasing by 5% one by one with the volumetric concentration of alcohol is a gradient, carry out wash-out, a gradient elution amount is a column volume, and every 10mL is collected in the test tube, point sample analysis on thin plate merges same composition; Each component is concentrated into moisture content and is no more than 5%, each component goes up the HW-40 post respectively, increasing by 5% one by one with the volumetric concentration of alcohol is a gradient, carry out wash-out, the employing Fraction Collector is collected, every 5mL is collected in the test tube, and the thin plate point sample merges same composition, obtains flavonoid compound monomer A, B, C, D, E, F, G.
(4) high performance liquid chromatography-mass spectrum, nuclear magnetic resonance technique carry out the structure evaluation to purified components: the flavonoid compound monomer that obtains is dissolved in the methyl alcohol, utilizes liquid matter logotype instrument to analyze.
Liquid-phase condition: chromatographic instrument: WATERS 2690; Detector: WATERS 996; Analytical column: Lichrospher C-18 2.1X250mm; Moving phase: methyl alcohol and 1% acetate gradient elution, methanol concentration from 0 to 100%; Column temperature: 30 ℃; Flow velocity: 0.3mL/min; Sample size: 10 μ L.
Mass spectrum condition: ionic means: EIS-, EIS+; Capillary voltage: EIS-is 3.88kV, and EIS+ is 3.87kV; Taper hole voltage: EIS-is 30V, and EIS+ is 24V; Ion source temperature: 120 ℃; Desolventizing temperature degree: 300 ℃; Mass range: 200-800m/z; Photomultiplier cell voltage: 650V; Analytical column is pressed: 2.6e-5mBar; Gas flow rate: 4.2L/hr.
Isolated flavonoid compound monomer is dissolved in the dimethyl sulfoxide (DMSO), on the Bruker nuclear magnetic resonance analyser, measures
1H-NMR,
13C-NMR, DEPT-NMR is an internal standard substance with the tetramethylsilane.
Beneficial effect of the present invention:
1, by AB-8 macroporous adsorbent resin, polyamide column and HW-40 post flavonoid compound in the Fructus Vaccinii Bracteati leaf melanochrome is carried out separation and purification, therefrom isolate 7 kinds of flavonoid compound monomers, be respectively A, B, C, D, E, F and G.
2, utilize sample length scanning collection of illustrative plates, HPLC-MS and nuclear magnetic resonance technique to carry out structural research, compd B, C, D, E and F have been carried out accurate structure to be identified, determine to be respectively Quercetin, chrysin, celery flavine, kaempferol and luteolin, its relative content is respectively: 37.51%, 2.26%, 9.57%, 1.72% and 15.16%.
Description of drawings
The length scanning collection of illustrative plates of Fig. 1 compd A
The HPLC collection of illustrative plates of Fig. 2 compd A
The MS collection of illustrative plates of Fig. 3 compd A
The MS collection of illustrative plates (positive ion) of Fig. 4 compd B
The MS collection of illustrative plates (negative ion) of Fig. 5 compd B
The length scanning collection of illustrative plates of Fig. 6 compound G
The HPLC collection of illustrative plates of Fig. 7 compound G
Specific embodiments
The structure of embodiment 1 compd A is identified
This material is further purified the comparison difficulty, so just carry out simple analysis because amount is less.This material ammonia cure presents yellow, carries out hydrochloric acid-zinc powder experiment, becomes redness, after leaving standstill red precipitate is arranged; Lead salt precipitation experimental result has tangible red precipitate; AlCl
3Experiment generates yellow and under UV-light fluorescence is arranged.This material is carried out length scanning, see Fig. 1, as can be seen from the figure, this compound has the characteristic absorbance of flavonoid compound.This material is carried out the HPLC purity testing, and concrete outcome is seen Fig. 2, and therefrom the purity of compd A is higher as can be seen, and as can be seen from Figure 3 this compound Flavone aglycone molecular weight is 302 in addition, and molecular weight 464 is speculated as and connects a glucose.Comprehensive above analysis, compd A may for a hydroxyl by the sugar-substituted Quercetin of a part grape.
The structure of embodiment 2 compd Bs is identified
This material is a pale yellow powder, is soluble in methyl alcohol, ethanol.Ammonia cure presents yellow, and institute thinks flavonoid compound.As can be seen from Figure 4 and Figure 5, the relative molecular mass of sample should be 302, and the relative molecular mass of this and Quercetin is consistent, can illustrate tentatively that the two is with a kind of material.The hydrogen spectrum (
1H-NMR): δ 7.67 (1H, H-2 '), δ 7.53 (1H, d, d, J=8.5Hz, 1.6Hz, H-6 '), δ 6.88 (1H, d, J=8.5Hz, H-5 '), δ 6.19 (1H, S, H-6), δ 6.41 (1H, S, H-8).
By the analysis to above collection of illustrative plates, and can to draw the compd B that extracts according to relevant reference be Quercetin.Molecular structural formula is as follows:
The structure of embodiment 3 Compound C is identified
This material is a pale yellow powder, and ammonia cure presents yellow, is soluble in methyl alcohol, ethanol.The hydrogen spectrum (
1H-NMR): δ 8.02 (2H, d, J=7.1, H-2 ', 6 '), δ 7.6 (1H, dd, J=7.1,6.8, H-4 '), δ 7.56 (2H, dd, J=7.1,6.8, H-3 ', 5 '), δ 6.9 (1H, S, H-3), and δ 6.5 (1H, d, J=2.0, H-8), δ 6.2 (1H, d, J=2.0, H-6).From
1H-NMR as can be seen, there are 8 hydrogen in the monocycle district, wherein two mutual couplings of hydrogen of δ 6.5 and δ 6.2, from the J value should monocycle between position hydrogen, show that this phenyl ring is 4 replacements.δ 8.02, and there are 5 hydrogen at δ 7.6 and δ 7.56 places, from peak shape and J value, should be a spin system, so be single-substituted ring.
The carbon spectrum (
13C-NMR): δ 181.8 (C4), δ 164.4 (C2), δ 163.1 (C8 '), δ 161.4 (C7), δ 157.4 (C5), δ 131.9 (C1 '), δ 130.7 (C4 '), δ 129.0 (C2 ', C6 '), δ 126.3 (C3 ', C5 '), δ 105.1 (C3), δ 103.9 (C4a), δ 99.0 (C8), δ 94.0 (C6).From
13As can be seen, this compound has 15 carbon on the C-NMR, and a carbonyl is arranged, and shows that this compound is a flavonoid compound.Therefrom comprise 4 even oxygen carbon in the analysis of compounds, illustrate except that the flavones parent nucleus, to also have two oxygen-containing substituents.In addition from
1There is a unimodal hydrogen in H-NMR at δ 6.9 places as can be seen, and this can illustrate that more this material is a flavonoid compound.Can judge that with reference to relevant document this compound is a chrysin, molecular structural formula is as follows:
The structure of embodiment 4 Compound D is identified
This material is the field gray powder, and ammonia cure presents yellow, is soluble in methyl alcohol, ethanol.The hydrogen spectrum (
1H-NMR): δ 7.91 (2H, d, J=8.6, H-2 ', 6 '), δ 6.93 (2H, d, J=8.7, H-3 ', 5 '), δ 6.75 (1H, s, H-3), δ 6.5 (1H, d, J=1.9, H-8), δ 6.19 (1H, d, J=1.9, H-6).From hydrogen spectrum (
1H-NMR) as can be seen, there are four hydrogen at δ 7.91 and δ 6.93 places, constitute AA ' BB ' system, so be four hydrogen on the disubstituted benzenes ring.There are two hydrogen at δ 6.5 and δ 6.19 places, constitute the AX system, and position hydrogen between being judged as by the J value is so have
Structural unit, the unimodal hydrogen meter at δ 6.75 places is bright to have only a hydrogen on a cyclic group in addition, so see that from the hydrogen spectrum this compound may be flavonoid compound.
The carbon spectrum (
13C-NMR): δ 181.7 (C4), δ 164.1 (C2), δ 163.7 (C8), δ 161.4 (C7), and δ 161.1 (C4 '), δ 157.3 (C5), δ 128.4 (C2 ', 6 '), δ 121.2 (C1 '), δ 115.9 (C3 ', 5 '), and δ 103.7 (C4a '), δ 102.8 (C3), δ 98.8 (C8), δ 93.9 (C6).From
13As can be seen, this compound has 15 carbon, and contains carbonyl on the C-NMR, can confirm clearly that this compound is a flavonoid compound.Also finding out from spectrum in addition has 5 even oxygen carbon, so also have three oxygen-containing substituents except that parent nucleus.Drawing this compound with reference to relevant document simultaneously from above data analysis is the celery flavine, and its structural formula is as follows:
The structure of embodiment 5 compd Es is identified
This material is a yellow powder, and ammonia cure presents yellow, is soluble in methyl alcohol, ethanol.The hydrogen spectrum (
1H-NMR): δ 8.06 (2H, d, J=8.8, H-2 ', 6 '), δ 6.94 (2H, d, J=8.8, H-3 ', 5 '), δ 6.4 (1H, d, J=1.8, H-8), δ 6.2 (1H, d, J=1.8, H-6).From hydrogen spectrum (
1H-NMR) at δ 8.06 and δ 6.94 places four hydrogen are arranged as can be seen, constitute AA ' BB ' system, showing has
Structure, there are two hydrogen at δ 6.4 and δ 6.2 places, constitute the AX system, and judging from the J value should be for a position hydrogen, so have
Structural unit.
The carbon spectrum (
13C-NMR): δ 175.9 (C4), δ 163.9 (C4 '), δ 160.7 (C8), δ 159.2 (C7), δ 156.2 (C5), δ 146.8 (C2), δ 135.6 (C3), δ 129.5 (C2 ', 6 '), δ 121.7 (C1), δ 115.4 (C3 ', 5 '), δ 103.0 (C4a), δ 98.2 (C8), δ 93.4 (C6).From
13As can be seen, this compound has 15 carbon on the C-NMR, and a carbonyl is arranged, and illustrates that this compound is a flavonoid compound.From the carbon of δ 135.6 as can be seen this compound be the flavonols compound.This compound has 6 even oxygen carbon as can be seen from spectrum, except that parent nucleus, also has four even oxygen substituting groups.By above analysis, and can know that this compound is a kaempferol with reference to relevant document, its molecular structural formula is as follows:
The structure of embodiment 6 compound F 17-hydroxy-corticosterones is identified
This material is the glassy yellow powder, and ammonia cure presents yellow, is soluble in methyl alcohol, ethanol.The hydrogen spectrum (
1H-NMR): δ 7.36 (2H, br, H-2 ', 5 '), δ 6.83 (1H, t, J=4.5, H-6 '), δ 6.67 (1H, d, J=2.0, H-8), δ 6.21 (1H, s, H-3), δ 6.08 (1H, d, J=2.0, H-6).From hydrogen spectrum (
1H-NMR) as can be seen, there are two hydrogen at δ 6.67 and δ 6.08 places, constitute the AX system, so have
Structural unit.There are three hydrogen at δ 7.36 and δ 6.83 places, see and have from peak shape and chemical potential in-migration
Structural unit.
The carbon spectrum (
13C-NMR): δ 181.2 (C4), δ 167.5 (C2), δ 163.6 (C8a), δ 161.4 (C7), δ 157.6 (C5), δ 151.3 (C4 '), δ 146.4 (C3 '), δ 120.5 (C1 '), δ 118.8 (C6 '), δ 116.0 (C5 '), δ 112.7 (C2 '), δ 102.5 (C4a), δ 102.1 (C3), δ 99.6 (C8), δ 94.3 (C6).From
13As can be seen, this compound has 15 carbon on the C-NMR, and a carbonyl is arranged, and illustrates that this compound is a flavonoid compound.6 even oxygen carbon are wherein arranged, except that parent nucleus, should also contain 4 even oxygen carbon.Can draw this compound by above analysis and the relevant document of reference is luteolin, and its molecular structural formula is as follows:
The structure of embodiment 7 compound G is identified
This material is owing to being further purified the comparison difficulty, so also just carry out simple analysis.This material ammonia cure presents yellow, carries out hydrochloric acid-zinc powder experiment, becomes redness, after leaving standstill red precipitate is arranged; Lead salt precipitation experimental result has tangible red precipitate; AlCl
3The colour developing experiment, generating yellow has a little yellow mercury oxide, and under UV-light fluorescence is arranged.This material is carried out length scanning, see Fig. 6, as can be seen from the figure, this compound has the characteristic absorbance of flavonoid compound.This material is carried out the HPLC purity testing, and concrete outcome is seen Fig. 7, and therefrom the purity of compound G is higher as can be seen.Comprehensive above analysis, compound G is a flavonoid compound.
Each compound relative content is measured
Each component relative content of table 1
| Compound | B | E | F | D | C |
| Appearance time (min) | 20.91 | 22.12 | 23.89 | 24.47 | 27.61 |
| Relative content (%) | 37.51 | 1.72 | 15.16 | 9.57 | 2.26 |
Remaining component awaits further research in the Fructus Vaccinii Bracteati leaf melanochrome extracting solution.
Claims (1)
1. the method identified of the monomeric extraction of flavonoid compound in the Fructus Vaccinii Bracteati leaf melanochrome, separation, purifying and structure, it is characterized in that with the Fructus Vaccinii Bracteati leaf be raw material, obtain Fructus Vaccinii Bracteati leaf melanochrome solution through extraction using alcohol, through macroporous adsorbent resin the flavonoid compound in the melanochrome is carried out roughing out again, by polyamide column and HW-40 post the flavonoid compound monomer is carried out purifying again, with high performance liquid chromatography-mass spectrum, nuclear magnetic resonance technique purified components is carried out structure again and identify;
(1) extraction of Fructus Vaccinii Bracteati leaf melanochrome solution: will clean up, air dried Fructus Vaccinii Bracteati leaf utilizes tissue mashing machine's wet method to smash to pieces, add reactor, utilize spirituous solution to extract, solid-liquid ratio is 1: 20～1: 30 in mass, the spirituous solution volumetric concentration is 60%, extracting temperature is 55～60 ℃, stir and extract 1h, solution is leached, obtain the aqueous solution of extract behind the recovered alcohol, the spirituous solution adjusted volume concentration that reclaims is to be added to reactor after 60%, to the leaf second extraction 1h in the reactor, solution is leached the aqueous solution merging that obtains the aqueous solution of extract behind the recovered alcohol and extract the extract that obtains for the first time according to the same terms, the moisture content that is concentrated into extracting solution is no more than 10%, and is standby;
(2) macroporous adsorbent resin carries out roughing out to the flavonoid compound in the melanochrome: add volume ratio under the extracting solution normal temperature and be 1: 1 n-butanol extraction half an hour, butanol extraction liquid is evaporated to does not have the alcohol flavor, the adding by volume is 1: 2～1: 3 a deionized water, sonic oscillation, AB-8 macroporous adsorptive resins on the aqueous solution, first deionized water wash-out with 2～3 times of column volumes, discard elutriant, be 95% alcohol wash-out again with the volumetric concentration of 3～5 times of column volumes, collect elutriant, be concentrated into ethanol content and be no more than 5%, standby;
(3) polyamide column and HW-40 post carry out purifying to the flavonoid compound monomer: the wash-out concentrated solution that macroporous adsorptive resins is eluted, last polymeric amide huge pillar, column volume φ 4cm * 50cm, the difference water, volumetric concentration is 20% alcohol, 45% alcohol, 70% alcohol, 95% alcohol carries out wash-out successively, each elutriant consumption is 2～3 column volumes, collect elutriant, point sample analysis on thin plate, merge identical component, same composition is concentrated into moisture content and is no more than 5%, and each component goes up the polymeric amide pillar respectively, φ 3cm * 30cm, increasing by 5% one by one with the volumetric concentration of alcohol is a gradient, carry out wash-out, a gradient elution amount is a column volume, and every 10mL is collected in the test tube, point sample analysis on thin plate merges same composition; Each component is concentrated into moisture content and is no more than 5%, each component goes up the HW-40 post respectively, increasing by 5% one by one with the volumetric concentration of alcohol is a gradient, carry out wash-out, the employing Fraction Collector is collected, every 5mL is collected in the test tube, and the thin plate point sample merges same composition, obtains flavonoid compound monomer A, B, C, D, E, F, G;
(4) high performance liquid chromatography-mass spectrum, nuclear magnetic resonance technique carry out the structure evaluation to purified components: the flavonoid compound monomer that obtains is dissolved in the methyl alcohol, utilizes liquid matter logotype instrument to analyze,
Liquid-phase condition: chromatographic instrument: WATERS 2690; Detector: WATERS 996; Analytical column: Lichrospher C-18 2.1 * 250mm; Moving phase: methyl alcohol and 1% acetate gradient elution, methanol concentration from 0 to 100%; Column temperature: 30 ℃; Flow velocity: 0.3mL/min; Sample size: 10 μ L;
Mass spectrum condition: ionic means: EIS-, EIS+; Capillary voltage: EIS-is 3.88kV, and EIS+ is 3.87kV; Taper hole voltage: EIS-is 30V, and EIS+ is 24V; Ion source temperature: 120 ℃; Desolventizing temperature degree: 300 ℃; Mass range: 200-800m/z; Photomultiplier cell voltage: 650V; Analytical column is pressed: 2.6e-5mBar; Gas flow rate: 4.2 L/hr;
Isolated flavonoid compound monomer is dissolved in the dimethyl sulfoxide (DMSO), on the Bruker nuclear magnetic resonance analyser, measures
1H-NMR,
13C-NMR, DEPT-NMR is an internal standard substance with the tetramethylsilane.
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