CN106501427A - The pre-treating method and simultaneous quantitative determination of multiple endogenous plant hormones in a kind of plant sample - Google Patents
The pre-treating method and simultaneous quantitative determination of multiple endogenous plant hormones in a kind of plant sample Download PDFInfo
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Abstract
The invention provides in a kind of plant sample multiple endogenous plant hormones pre-treating method and quantitative detecting method, in plant sample, 1) be quantitatively adding the Isotopic Internal Standard of multiple endogenous plant hormones, sample solution obtained with solvent extraction then；2) it is purification medium to step 1 with graphitized charcoal black-materials) gained sample solution carries out remove impurity, and then obtain the solution to be measured of multiple endogenous plant hormones in plant sample；3) solution to be measured of multiple endogenous plant hormones in gained plant sample is gathered response data by analytical tool, and then using the multiple endogenous plant hormones in internal standard method Simultaneous Determination plant sample.The inventive method is simple, quick, province's solvent, easy to operate, the enriching and purifying of the endogenous plant hormone of achievable low content, and realizes that simultaneous quantitative is detected, method is simple and quick, and accuracy is high.
Description
Technical field
The present invention relates to the pre-treating method of multiple endogenous plant hormones and simultaneous quantitative detection in a kind of plant sample
Method, belongs to analytical chemistry field.
Background technology
Phytohormone was played and extremely important effect in each stage of plant growing, development.Simultaneously, accurately fixed
Various plants hormone in amount plant tissue, contributes to physiological function and the regulated and control network for understanding phytohormone.But, due to plant
Extracting solution mesostroma is complicated, content is extremely low and differing chemical properties so that while determining various plants hormone tool in plant sample
There is larger difficulty.
In the plant that has reported at present, the pre-treating method of various plants hormone includes liquid-liquid extraction, Solid-Phase Extraction, solid phase
Micro-extraction, magnetic solid phase extraction and liquid chromatography purification method etc., these method pretreatment process majorities can bring solvent consumption big
And the problems such as high cost.Further, since chemical property differs greatly, above-mentioned pre-treating method can inevitably result in the response rate
Low, detection sensitivity is poor, the detection being unfavorable in actual sample.
Existing patent ZL 2,012 1 0486507.3 and ZL 2,013 1 0131422.8, are based on Brassica campestris L element sterol
(BRs) analysis detection.As BRs parahormone chemical constitution properties are similar, and all contain cis hydroxyl groups, for this class material
Property, the pre-treatment scheme having described in above-mentioned patent.But for chemical property is different from its of Brassica campestris L element sterol (BRs)
His class phytohormone, cannot then be realized with such scheme.
Content of the invention
The technical problem to be solved is the deficiency and a kind of plant sample of offer existed for above-mentioned prior art
The pre-treating method and simultaneous quantitative determination of multiple endogenous plant hormones in product, can realize the multiple interior of low content simultaneously
The enriching and purifying of source property phytohormone, and realize that simultaneous quantitative is detected, method is simple and quick, and accuracy is high.
The present invention by solve the problems, such as the technical scheme for adopting set forth above for：
The pre-treating method of multiple endogenous plant hormones in a kind of plant sample, key step are as follows：
1) Isotopic Internal Standard of multiple endogenous plant hormones is quantitatively adding in plant sample, is then obtained with solvent extraction
Arrive sample solution；
2) it is purification medium to step 1 with graphitized charcoal black-materials) gained sample solution carries out remove impurity, and then obtain plant
The solution to be measured of multiple endogenous plant hormones in sample, so that realize the sample of multiple endogenous plant hormones in plant sample
Pre-treatment.
Press such scheme, Oryza sativa L. of the plant sample for Seedling Stage, the Oryza sativa L. in tillering stage, the blade and flower of pustulation period
Fringe, the blade and spica in period of maturation, and the root of Oryza sativa L., the root of arabidopsiss, stem, leaf, flower and silique etc. plant sample.
Such scheme is pressed, the plant sample is powder, such as can in advance using being ground to powder after liquid nitrogen freezing.
Press such scheme, the species of the endogenous plant hormone mainly includes auxins, cytokinin, red mould
Endogenous plant hormone of plain class, Jasmonates, salicylic acid and the acids that comes off etc..
Such scheme is pressed, the endogenous plant hormone mainly includes indole-3-acetic acid (IAA), indole -3-butyric acid
(IBA), abscisic acid (ABA), salicylic acid (SA), trans-zeatin (tZ), trans-zeatin nucleoside (tZR), cis zeatin
(cZ), cis ribosylzeatin (cZR), isopentenyl gland purine (iP), isopentenyl gland purine nucleoside (iPR), double hydrogen Semen Maydiss
Plain (DZ), double hydrogen ribosylzeatin (DZR), trans-zeatin -7- glucosides (tZ7G), trans-zeatin -9- glucosides (tZ9G) are suitable
Formula zeatin -9- glucosides (cZ9G), cis zeatin-O-glycosides (cZOG), double hydrogen zeatin -7- glucosides (DZ7G), double hydrogen are beautiful
Meter Su -9- glucosides (DZ9G), double hydrogen zeatin-O-glycosides (DZOG), isopentenyl gland purine -7- glucosides (iP7G), iso-amylene
Base adenine -9- glucosides (iP9G), 2- benzylthio isopentenyl gland purines (2BSiP), 2- chlorine trans-zeatins (2CltZ), 2-
Methyl mercapto trans-zeatin (2MeStZ), the cis zeatin of 2- methyl mercaptos (2MeScZ), 2- methyl mercapto trans-zeatin nucleoside
(2MeStZR), the cis ribosylzeatin of 2- methyl mercaptos (2MeScZR), 2- methyl mercapto isopentenyl gland purines (2MeSiP), 2- first
Sulfenyl isopentenyl gland purine nucleoside (2MeSiPR), 1 (GA of gibberellins1), 3 (GA of gibberellins3), 4 (GA of gibberellins4), gibberellins 5
(GA5), 6 (GA of gibberellins6), 7 (GA of gibberellins7), 8 (GA of gibberellins8), 9 (GA of gibberellins9), 12 (GA of gibberellins12), gibberellins
13(GA13), 15 (GA of gibberellins15), 19 (GA of gibberellins19), 20 (GA of gibberellins20), 23 (GA of gibberellins23), gibberellins 24
(GA24), 29 (GA of gibberellins29), 34 (GA of gibberellins34), 44 (GA of gibberellins44), 51 (GA of gibberellins51), gibberellins 53
(GA53), jasmonic (JA), jasmonic-isoleucine (JA-leu), jasmonic-phenylalanine (JA-phe), 12- hydroxyl jasmines
Sour (12OHJA), 12- oxos phytadiene acid (OPDA), one or more in abscisic acid (ABA) and salicylic acid (SA) etc..
Such scheme is pressed, corresponding Isotopic Internal Standard is added as needed, predominantly [2H2] IAA, [2H6] ABA, dihydro jasmine
Jasmine acid (2H-JA), [2H4] SA, [2H2]GA1, [2H2]GA4, [2H2]GA5, [2H2]GA6, [2H2]GA7, [2H2]GA8, [2H2]GA9,
[2H2]GA12, [2H2]GA15, [2H2]GA20, [2H2]GA24, [2H2]GA34, [2H2]GA44, [2H2]GA51, [2H2]GA53, [2H5] tZ,
[15N5] cZ, [2H5] tZ7G, [2H5] tZ9G, [2H3] DZ, [2H5] DZ9G, [2H7] DZOG, [2H6] iP, [2H6] iP9G, [2H5]
TZR, [2H3] DZR, [2H6] iPR etc..The corresponding internal standard of multiple endogenous plant hormones to be measured is as shown in table 1.
Press such scheme, the solvent be methanol, acetonitrile, isopropanol, chloroform, dichloromethane, ethyl acetate, normal hexane,
One or more in ether, acetone or formic acid etc. mixture in any proportion.The effect of solvent is：Efficiently extracting out
On the premise of analyte, the chaff interferences such as impurity are extracted as few as possible.
Such scheme is pressed, the temperature of the solvent extraction is -25～-15 DEG C, and time range is 6～12h.
Such scheme is pressed, the temperature of remove impurity is room temperature after the addition graphitized charcoal black-materials, and the time is 3min or big
In 3min.The mode such as vortex or vibration can be adopted uniformly abundant to ensure remove impurity in dedoping step.
Press such scheme, step 2) in preferably step be：First, removing step solvent 1) in gained sample solution,
Add graphitized charcoal black-materials to carry out remove impurity after redissolution, collect gained supernatant after remove impurity, multiple endogenous as in plant sample
The solution to be measured of property phytohormone.
Press such scheme, step 2) in more preferably step be：First, removing step is 1) molten in gained sample solution
Agent, obtains sample slag；Then gained sample slag adds solvent B to redissolve, the sample solution after being redissolved；Then, gained is multiple
Add graphitized charcoal black-materials to carry out remove impurity in sample solution after molten, collect gained supernatant after remove impurity；Finally, after by remove impurity
Solution B in gained supernatant is removed, and adds solvent C to redissolve, and obtains the to be measured of multiple endogenous plant hormones in plant sample
Solution.Wherein, solvent B is not for sample dissolution slag, and can adsorbing or the multiple endogenous plants of few adsorbed target analyte swash
The solvent of element, such as methanol and its mixed solution with water, acetonitrile and its with the mixed solution of water etc.；The solvent C can with rear
Continuous separation condition is adapted, such as methanol and its mixed solution with water, acetonitrile and its with the mixed solution of water etc..
Press such scheme, the step 2) in graphitized charcoal black-materials be the charcoal base granule containing π-pi-conjugated group and
One or more in gel, magnetic material, amorphous material, packed column or integral post comprising this kind of charcoal base granule etc..Institute
The remove impurity that states refers to and removes a large amount of pigments, lipid and the strong-hydrophobicity impurity that contain in sample solution, thus adopt by material with carbon element for
Substrate, the silica gel particle of structure, packed column or integral post etc. can reach the purpose of remove impurity.
The present invention also provides multiple endogenous plant hormones in a kind of plant sample on the basis of above-mentioned pre-treating method
While quantitative detecting method, the solution to be measured of multiple endogenous plant hormones in pre-treating method gained plant sample is passed through
Analytical tool collection response signal or data, and then using the multiple endogenouss in internal standard method Simultaneous Determination plant sample
Phytohormone.
Such scheme is pressed, the analytical tool can adopt liquid chromatograph-fluorescence detector, liquid chromatograph-mass spectrometer to join
With, capillary electrophoresis-fluorescence detector combination, the combination of capillary electrophoresis-mass spectrometry instrument etc..Preferably, the analytical tool is adopted
High performance liquid chromatography-electron spray-triple quadrupole rods tandem mass spectrometries.
Specifically, quantitative detecting method while multiple endogenous plant hormones in a kind of plant sample, key step is such as
Under：
1) the standard sample solution of variable concentrations gradient is configured, and is quantitatively adding the isotope of multiple endogenous plant hormones
Internal standard, then carries out Ultra Performance Liquid Chromatography-tandem mass spectrum (UPLC-MS/MS) analysis to configured good standard solution and obtains
The chromatogram peak area of each material (i.e. every kind of endogenous plant hormone) is then integrated, divided by corresponding by chromatogram
Interior target peak area, makees linear regression to the concentration of respective substance (i.e. every kind of endogenous plant hormone), and standard curve is obtained；
2) Isotopic Internal Standard of multiple endogenous plant hormones is quantitatively adding in plant sample, is then obtained with solvent extraction
Arrive sample solution；
3) it is purification medium to step 2 with graphitized charcoal black-materials) gained sample solution carries out remove impurity, and then obtain plant
The solution to be measured of multiple endogenous plant hormones in sample；
4) solution to be measured of multiple endogenous plant hormones in gained plant sample is passed through Ultra Performance Liquid Chromatography-series connection
Mass spectrum (UPLC-MS/MS) analysis collection chromatogram；Further by each material in chromatogram (i.e. every kind of endogenous plant hormone)
Integrating peak areas, divided by corresponding interior target peak area, using step 1) gained standard curve Simultaneous Determination plant
Multiple endogenous plant hormones in sample.
Compared with prior art, the invention has the beneficial effects as follows：
1st, the inventive method is simple, quick, save solvent, easy to operate, can achieve the richness of the endogenous plant hormone of low content
Collection purification, and detection by quantitative while multiple endogenous plant hormones in plant sample is realized, method is simple and quick, accuracy
High.
2nd, compared with the prior art being previously mentioned in background technology, the present invention can realize up to 54 kinds and above simultaneously
The analysis detection of distinct phytohormone, rather than it is only applicable to a class phytohormone (Brassica campestris L element sterol) detection.
3rd, compared with the prior art being previously mentioned in background technology, in the present invention, Graphon can realize plant extract
The purification of liquid, and not adsorbed target analyte --- distinct phytohormone, achieve unexpected technique effect；And
It is previously mentioned in background technology in patent ZL 2,012 1 0486507.3, Graphon is only used as a part for pre-treatment, scheme
Complexity, and adsorb more non-Study on Brassinosteroids.
4th, the present invention can adopt the scheme of a step pre-treatment, reduce loss of the target analytes in front process；Meanwhile,
Using more efficient ultra high efficiency liquid phase systems are separated, numerous molecular weight identical compounds can be separated, 54 is achieved with this
The detection of Plant Hormone.
Description of the drawings
Fig. 1 is many reaction monitoring chromatograms of 54 Plant Hormones that the inventive method can be implemented to analyze.Wherein, (A),
peak 1-54；(B),peak1-7；(C),peak22-27；(D),peak38-42；1,tZ7G；2,tZ；3,DZ；4,cZOG；5,
DZ7G；6,DZOG；7,cZ；8,DZ9G；9,tZ9G；10,cZ9G；11,iP7G；12,iP；13,DZR；14,tZR；15,GA8；16,
cZR；17,GA29；18,iP9G；19,12OHJA；20,GA23；21,SA；22,2CltZ；23,GA3；24,iPR；25,2MeStZ；
26,2MeScZ；27,GA1；28,IAA；29,GA6；30,2MeStZR；31,2MeStZR；32,ABA；33,GA13；34,GA5；35,
GA19；36,GA20；37,GA44；38,JA；39,IBA；40,GA34；41,2MeSiP；42,GA51；43,GA53；44,2MeSiPR；
45,GA7；46,GA4；47,GA24；48,JA-leu；49,JA-phe；50,GA15；51,GA9；52,2BSiP；53,GA12；54,
OPDA.
Fig. 2 is the content of endogenous plant hormone in the different plant tissues that method of the present invention is detected.Wherein (A) is plant
Distribution thermal map of the hormone in Oryza sativa L. sample；(B) cell divide element and total content of the gibberellins in Oryza sativa L. and dissimilar
Basic element of cell division proportion.
Specific embodiment
For a better understanding of the present invention, with reference to embodiment further to the present invention provided technical scheme in
Key technology is further described, but the present invention is not limited solely to the following examples.
Embodiment 1
The pre-treating method of multiple endogenous plant hormones in a kind of plant sample, key step are as follows：
(1) accurately weigh 50mg plant tissues (the parallel Oryza sativa L. for taking Seedling Stage respectively, the Oryza sativa L. in tillering stage, the pustulation period
The plant tissue of root these different parts of blade and spica, the blade and spica in period of maturation, and Oryza sativa L. as pre-treatment with
And the plant sample of subsequent quantitation detection), it is transferred in 1.5mL centrifuge tubes, is quantitatively adding the same of multiple endogenous plant hormones
The plain internal standard in position [2H2]IAA(1.0ng),[2H6]ABA(1.0ng),2H-JA(1.0ng)[2H4]SA(50ng),[2H2]GA1
(0.5ng),[2H2]GA4(0.5ng),[2H2]GA5(0.5ng),[2H2]GA6(0.5ng),[2H2]GA7(0.5ng),[2H2]GA8
(0.5ng),[2H2]GA9(0.5ng),[2H2]GA12(0.5ng),[2H2]GA15(0.5ng),[2H2]GA20(0.5ng),[2H2]
GA24(0.5ng),[2H2]GA34(0.5ng),[2H2]GA44(0.5ng),[2H2]GA51(0.5ng),[2H2]GA53(0.5ng),
[2H5]tZ(0.1ng),[2H5]cZ(0.1ng),[2H5]tZ7G(0.1ng),[2H5]tZ9G(0.1ng),[2H3]DZ(0.1ng),
[2H5]DZ9G(0.1ng),[2H7]DZOG(0.1ng),[2H6]iP(0.1ng),[2H6]iP9G(0.1ng),[2H5]tZR
(0.1ng),[2H3]DZR(0.1ng),[2H6] iPR (0.1ng), place in a moment, add 0.5mL acetonitrile mix homogeneously, be put into-
Extract 12 hours in 20 DEG C of refrigerator, centrifuging and taking supernatant, as sample solution；
(2) solvent in removing step (1) gained sample solution, is redissolved with 80% (v/v) acetonitrile solution, and gained is multiple
Sample solution after molten is added in the centrifuge tube of the 1.5mL containing 10mg Graphons, and being vortexed carries out remove impurity in 3 minutes, from
The heart takes supernatant and dries up to remove solvent, then is redissolved with 100 microlitre 5% of acetonitrile solution, that is, obtain many in plant sample
The solution to be measured of endogenous plant hormone is planted, so as to realize the sample pre-treatments of multiple endogenous plant hormones in plant sample.
Quantitative detecting method while multiple endogenous plant hormones in a kind of plant sample, key step are as follows：
1) configure variable concentrations gradient standard sample solution (concentration range is as shown in table 1), and add internal standard [2H2]IAA
(1.0ng),[2H6]ABA(1.0ng),2H-JA(1.0ng)[2H4]SA(50ng),[2H2]GA1(0.5ng),[2H2]GA4
(0.5ng),[2H2]GA5(0.5ng),[2H2]GA6(0.5ng),[2H2]GA7(0.5ng),[2H2]GA8(0.5ng),[2H2]GA9
(0.5ng),[2H2]GA12(0.5ng),[2H2]GA15(0.5ng),[2H2]GA20(0.5ng),[2H2]GA24(0.5ng),[2H2]
GA34(0.5ng),[2H2]GA44(0.5ng),[2H2]GA51(0.5ng),[2H2]GA53(0.5ng),[2H5]tZ(0.1ng),[2H5]
cZ(0.1ng),[2H5]tZ7G(0.1ng),[2H5]tZ9G(0.1ng),[2H3]DZ(0.1ng),[2H5]DZ9G(0.1ng),
[2H7]DZOG(0.1ng),[2H6]iP(0.1ng),[2H6]iP9G(0.1ng),[2H5]tZR(0.1ng),[2H3]DZR
(0.1ng),[2H6]iPR(0.1ng)；
2) to step 1) standard solution that configured carries out Ultra Performance Liquid Chromatography-tandem mass spectrum (UPLC-MS/MS) point
Analysis, obtains chromatogram as shown in Figure 1, then integrates the chromatogram peak area of each material, divided by corresponding interior target
Peak area, makees linear regression to the concentration of respective substance, and standard curve (as shown in table 1) is obtained；
3) by the solution to be measured of the phytohormone at the present embodiment pre-treating method gained difference rice tissue position by super
High performance liquid chromatography-tandem mass (UPLC-MS/MS) analysis collection chromatogram；Further by the peak face of each material in chromatogram
Product integration, divided by corresponding interior target peak area, using step 2) gained standard curve Simultaneous Determination plant sample
In multiple endogenous plant hormones, the plant hormone levels for measuring different rice tissue positions are as shown in Figure 1.
The working curve of 1 analyte of table
For the accuracy of verification method, the present invention adds the basic element of cell division mark product of variable concentrations in Oryza sativa L. sample
(basic, normal, high, refer to table 2), processed according to described method afterwards and determined, draw the response rate of different spiked levels
Between 80.3%-120.4%, show the accuracy of method provided by the present invention.In order to investigate the precision of method, at one day
The interior sample to different spiked levels carries out 5 times to be repeated to process and determines, and calculates the relative standard deviation of the respective response rate
(RSD), as withinday precision；Continuous three days samples to different spiked levels are processed and are determined, and are calculated respective time
The relative standard deviation (RSD) of yield, as day to day precision.As a result as shown in table 2, as a result RSD shows this less than 11.8%
Invention methods described is reliable and stable.
The degree of stability of 2 method of table and accuracy
In sum, the present invention can adopt the scheme of a step pre-treatment, can achieve the endogenous plant hormone of low content
Enriching and purifying, simple, quick, save solvent, easy to operate, and subsequently can achieve up to 54 kinds and above distinct plant swashs
Detection by quantitative while plain, method are simple and quick, and accuracy is high, have jumped out in prior art a kind of pre-treatment and its quantitative completely
Detection method is only capable of the limitation for a class phytohormone.
The above is only the preferred embodiment of the present invention, it is noted that for one of ordinary skill in the art comes
Say, without departing from the concept of the premise of the invention, some modifications and variations can also be made, these belong to the present invention's
Protection domain.
Claims (10)
1. in a kind of plant sample multiple endogenous plant hormones pre-treating method, it is characterised in that key step is as follows：
1) Isotopic Internal Standard of multiple endogenous plant hormones is quantitatively adding in plant sample, and sample is obtained with solvent extraction then
Product solution；
2) it is purification medium to step 1 with graphitized charcoal black-materials) gained sample solution carries out remove impurity, and then obtain plant sample
In multiple endogenous plant hormones solution to be measured, so as to realize in plant sample locating before the sample of multiple endogenous plant hormones
Reason.
2. in a kind of plant sample according to claim 1 multiple endogenous plant hormones pre-treating method, its feature
Be the endogenous plant hormone species mainly include auxins, cytokinin, gibberellins class, Jasmonates,
Salicylic acid and the endogenous plant hormone of the acids that comes off.
3. in a kind of plant sample according to claim 2 multiple endogenous plant hormones pre-treating method, its feature
It is that the endogenous plant hormone mainly includes indole-3-acetic acid, indole -3-butyric acid, abscisic acid, salicylic acid, trans Semen Maydiss
Element, trans-zeatin nucleoside, cis zeatin, cis ribosylzeatin, isopentenyl gland purine, isopentenyl gland purine core
Glycosides, double hydrogen zeatin, double hydrogen ribosylzeatins, trans-zeatin -7- glucosides, trans-zeatin -9- glucosides, cis zeatin -
9- glucosides, cis zeatin-O-glycosides, double hydrogen zeatin -7- glucosides, double hydrogen zeatin -9- glucosides, double hydrogen zeatin-O- sugar
Glycosides, isopentenyl gland purine -7- glucosides, isopentenyl gland purine -9- glucosides, 2- benzylthio isopentenyl gland purines, 2- chlorine are anti-
Formula zeatin, 2- methyl mercapto trans-zeatins, the cis zeatin of 2- methyl mercaptos, 2- methyl mercapto trans-zeatin nucleoside, 2- first sulfur
The cis ribosylzeatin of base, 2- methyl mercapto isopentenyl gland purines, 2- methyl mercapto isopentenyl gland purine nucleoside, gibberellins 1 are red
Mycin 3, gibberellins 4, gibberellins 5, gibberellins 6, gibberellins 7, gibberellins 8, gibberellins 9, gibberellins 12, gibberellins 13 are red mould
Element 15, gibberellins 19, gibberellins 20, gibberellins 23, gibberellins 24, gibberellins 29, gibberellins 34, gibberellins 44, gibberellins 51,
Gibberellins 53, jasmonic, jasmonic-isoleucine, jasmonic-phenylalanine, 12- hydroxyl jasmonics, 12- oxo phytadienes
Acid, abscisic acid and salicylic acid.
4. in a kind of plant sample according to claim 1 multiple endogenous plant hormones pre-treating method, its feature
It is that the solvent is methanol, acetonitrile, isopropanol, chloroform, dichloromethane, ethyl acetate, normal hexane, ether, acetone or formic acid
One or more in mixture in any proportion.
5. in a kind of plant sample according to claim 1 multiple endogenous plant hormones pre-treating method, its feature
Be the solvent extraction temperature be -25～-15 DEG C, time range be 6～12h.
6. in a kind of plant sample according to claim 1 multiple endogenous plant hormones pre-treating method, its feature
It is more than 3min that miscellaneous time is removed after being the addition graphitized charcoal black-materials.
7. in a kind of plant sample according to claim 1 multiple endogenous plant hormones pre-treating method, its feature
Be step 2) in concretely comprise the following steps：First, removing step solvent 1) in gained sample solution, adds graphitized charcoal after redissolution
Black-materials carry out remove impurity, collect gained supernatant after remove impurity, and as in plant sample, multiple endogenous plant hormones is to be measured molten
Liquid.
8. in a kind of plant sample according to claim 1 multiple endogenous plant hormones pre-treating method, its feature
Be the step 2) in graphitized charcoal black-materials be charcoal base granule and its magnetic material containing π-pi-conjugated group, without fixed
One or more in section bar material, gel, packed column or integral post.
9. quantitative detecting method while multiple endogenous plant hormones in a kind of plant sample, it is characterised in that key step is such as
Under：
1) Isotopic Internal Standard of multiple endogenous plant hormones is quantitatively adding in plant sample, and sample is obtained with solvent extraction then
Product solution；
2) it is purification medium to step 1 with graphitized charcoal black-materials) gained sample solution carries out remove impurity, and then obtain plant sample
In multiple endogenous plant hormones solution to be measured；
3) solution to be measured of multiple endogenous plant hormones in gained plant sample is gathered response data by analytical tool, is entered
And adopt the multiple endogenous plant hormones in internal standard method Simultaneous Determination plant sample.
10. quantitative detecting method while multiple endogenous plant hormones in a kind of plant sample according to claim 9,
It is characterized in that the analytical tool selected from liquid chromatograph-fluorescence detector, liquid chromatograph-mass spectrometer combination, capillary electrophoresis-
In fluorescence detector combination, the combination of capillary electrophoresis-mass spectrometry instrument.
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| CN107367556A (en) * | 2017-06-30 | 2017-11-21 | 山东农业大学 | Method that is a kind of while detecting seven kinds of endogenous hormones in wheat leaf blade |
| CN107796893A (en) * | 2017-08-31 | 2018-03-13 | 四川农业大学 | HPLC methods that are a kind of while determining 8 kinds of Endogenous Hormone Contents in Vitro in Paeonia lactiflora seed |
| CN108107131A (en) * | 2017-12-23 | 2018-06-01 | 中国农业科学院农业资源与农业区划研究所 | The method that soil various plants hormone can be detected simultaneously |
| CN108535374A (en) * | 2018-03-23 | 2018-09-14 | 山东农业大学 | A kind of method of auxin in measurement plant tissue |
| CN109324139A (en) * | 2018-12-14 | 2019-02-12 | 广西中烟工业有限责任公司 | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
| CN109324140A (en) * | 2018-12-14 | 2019-02-12 | 广西中烟工业有限责任公司 | Ribosylzeatin Solid Phase Extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
| CN109374763A (en) * | 2018-10-18 | 2019-02-22 | 中国科学院遗传与发育生物学研究所 | The quantitative detecting method of plant endo phytosulfokine-α content |
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| CN111308006A (en) * | 2020-03-16 | 2020-06-19 | 上海中科新生命生物科技有限公司 | LC-MS-based high-throughput high-sensitivity phytohormone detection method |
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| CN107367556A (en) * | 2017-06-30 | 2017-11-21 | 山东农业大学 | Method that is a kind of while detecting seven kinds of endogenous hormones in wheat leaf blade |
| CN107367556B (en) * | 2017-06-30 | 2019-07-30 | 山东农业大学 | Method that is a kind of while detecting seven kinds of endogenous hormones in wheat leaf blade |
| CN107796893A (en) * | 2017-08-31 | 2018-03-13 | 四川农业大学 | HPLC methods that are a kind of while determining 8 kinds of Endogenous Hormone Contents in Vitro in Paeonia lactiflora seed |
| CN108107131A (en) * | 2017-12-23 | 2018-06-01 | 中国农业科学院农业资源与农业区划研究所 | The method that soil various plants hormone can be detected simultaneously |
| CN108107131B (en) * | 2017-12-23 | 2020-07-03 | 中国农业科学院农业资源与农业区划研究所 | Method for simultaneously detecting multiple phytohormones in soil |
| CN108535374A (en) * | 2018-03-23 | 2018-09-14 | 山东农业大学 | A kind of method of auxin in measurement plant tissue |
| CN109374763A (en) * | 2018-10-18 | 2019-02-22 | 中国科学院遗传与发育生物学研究所 | The quantitative detecting method of plant endo phytosulfokine-α content |
| CN109324140A (en) * | 2018-12-14 | 2019-02-12 | 广西中烟工业有限责任公司 | Ribosylzeatin Solid Phase Extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
| CN109324139A (en) * | 2018-12-14 | 2019-02-12 | 广西中烟工业有限责任公司 | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
| CN109991339A (en) * | 2019-05-09 | 2019-07-09 | 中国科学院化学研究所 | A kind of sample pretreating method measuring endogenous hormones in trace plant sample |
| CN111044627A (en) * | 2019-12-03 | 2020-04-21 | 贵州民族大学 | Method for detecting endogenous hormones in pinus massoniana branches |
| CN111308006A (en) * | 2020-03-16 | 2020-06-19 | 上海中科新生命生物科技有限公司 | LC-MS-based high-throughput high-sensitivity phytohormone detection method |
| CN111308006B (en) * | 2020-03-16 | 2022-12-13 | 上海中科新生命生物科技有限公司 | LC-MS-based high-throughput high-sensitivity phytohormone detection method |
| CN112229928A (en) * | 2020-10-06 | 2021-01-15 | 武汉迈特维尔生物科技有限公司 | Method for simultaneously quantifying multiple phytohormones in plant sample |
| CN112229928B (en) * | 2020-10-06 | 2021-09-14 | 武汉迈特维尔生物科技有限公司 | Method for simultaneously quantifying multiple phytohormones in plant sample |
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