CN118209654B - A method for simultaneous detection of four cytokinins in litchi using UPLC-MS/MS - Google Patents
A method for simultaneous detection of four cytokinins in litchi using UPLC-MS/MS Download PDFInfo
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- UQHKFADEQIVWID-UHFFFAOYSA-N cytokinin Natural products C1=NC=2C(NCC=C(CO)C)=NC=NC=2N1C1CC(O)C(CO)O1 UQHKFADEQIVWID-UHFFFAOYSA-N 0.000 title claims abstract description 37
- 239000004062 cytokinin Substances 0.000 title claims abstract description 37
- 241001629511 Litchi Species 0.000 title claims abstract description 32
- 238000001514 detection method Methods 0.000 title claims abstract description 29
- 238000010811 Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry Methods 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 23
- 238000004458 analytical method Methods 0.000 claims abstract description 15
- 238000010828 elution Methods 0.000 claims abstract description 12
- 238000004949 mass spectrometry Methods 0.000 claims abstract description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 73
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 39
- 239000000243 solution Substances 0.000 claims description 32
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 22
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 18
- 239000007864 aqueous solution Substances 0.000 claims description 15
- 239000003480 eluent Substances 0.000 claims description 15
- 150000002500 ions Chemical class 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 13
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 11
- 235000019253 formic acid Nutrition 0.000 claims description 11
- 239000007789 gas Substances 0.000 claims description 10
- 239000012071 phase Substances 0.000 claims description 10
- 239000006228 supernatant Substances 0.000 claims description 10
- 229910052757 nitrogen Inorganic materials 0.000 claims description 9
- XKRFYHLGVUSROY-UHFFFAOYSA-N Argon Chemical compound [Ar] XKRFYHLGVUSROY-UHFFFAOYSA-N 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 7
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 7
- 239000007791 liquid phase Substances 0.000 claims description 7
- 238000000746 purification Methods 0.000 claims description 6
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical compound [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 claims description 5
- 238000000605 extraction Methods 0.000 claims description 5
- 229910052786 argon Inorganic materials 0.000 claims description 4
- 238000004807 desolvation Methods 0.000 claims description 4
- 238000000227 grinding Methods 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- 239000000284 extract Substances 0.000 claims description 3
- 239000006199 nebulizer Substances 0.000 claims 1
- 238000004128 high performance liquid chromatography Methods 0.000 abstract description 11
- 230000035945 sensitivity Effects 0.000 abstract description 8
- 238000000926 separation method Methods 0.000 abstract description 7
- 150000001875 compounds Chemical class 0.000 abstract description 6
- 238000001195 ultra high performance liquid chromatography Methods 0.000 abstract description 6
- 230000007547 defect Effects 0.000 abstract description 4
- 239000003375 plant hormone Substances 0.000 abstract 1
- HYVABZIGRDEKCD-UHFFFAOYSA-N N(6)-dimethylallyladenine Chemical compound CC(C)=CCNC1=NC=NC2=C1N=CN2 HYVABZIGRDEKCD-UHFFFAOYSA-N 0.000 description 8
- 238000002386 leaching Methods 0.000 description 6
- 238000001819 mass spectrum Methods 0.000 description 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 230000003213 activating effect Effects 0.000 description 3
- 238000007664 blowing Methods 0.000 description 3
- 239000000287 crude extract Substances 0.000 description 3
- 239000004047 hole gas Substances 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000003595 spectral effect Effects 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000002777 nucleoside Substances 0.000 description 2
- XXFACTAYGKKOQB-UHFFFAOYSA-N 2-methyl-4-(7h-purin-6-ylamino)butan-1-ol Chemical compound OCC(C)CCNC1=NC=NC2=C1NC=N2 XXFACTAYGKKOQB-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- XXFACTAYGKKOQB-SSDOTTSWSA-N Dihydrozeatin Natural products OC[C@H](C)CCNC1=NC=NC2=C1NC=N2 XXFACTAYGKKOQB-SSDOTTSWSA-N 0.000 description 1
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- GHQPBDDZGPAVJP-UHFFFAOYSA-N azanium;methanol;hydroxide Chemical compound N.O.OC GHQPBDDZGPAVJP-UHFFFAOYSA-N 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- XXFACTAYGKKOQB-ZETCQYMHSA-N dihydrozeatin Chemical compound OC[C@@H](C)CCNC1=NC=NC2=C1NC=N2 XXFACTAYGKKOQB-ZETCQYMHSA-N 0.000 description 1
- 230000005672 electromagnetic field Effects 0.000 description 1
- 238000011067 equilibration Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 1
- 229930195732 phytohormone Natural products 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- UZKQTCBAMSWPJD-UQCOIBPSSA-N trans-Zeatin Natural products OCC(/C)=C\CNC1=NC=NC2=C1N=CN2 UZKQTCBAMSWPJD-UQCOIBPSSA-N 0.000 description 1
- -1 trans-zeatin nucleoside Chemical class 0.000 description 1
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
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- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G01N30/8675—Evaluation, i.e. decoding of the signal into analytical information
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Abstract
本发明涉及植物激素技术领域，具体涉及一种用UPLC‑MS/MS同时检测荔枝中四种细胞分裂素的方法。该方法采用了超高效液相色谱(UPLC)，UPLC在HPLC的基础上进一步地提高了分离度和分析速度，质谱MS根据原子或分子的质荷比，将原子或分子电离，从而实现化合物的分离和检测，具有专属鉴定能力，MS具有高选择性、高灵敏度的特点，有效地弥补了HPLC选择性与灵敏度不够的缺陷，避免HPLC由于共流出导致不能有效地分离结构和极性相似的多种化合物的问题，为后续能够同时检测出荔枝中四种结构相近的细胞分裂素打下基础。
The present invention relates to the technical field of plant hormones, and in particular to a method for simultaneously detecting four cytokinins in litchi using UPLC-MS/MS. The method adopts ultra-high performance liquid chromatography (UPLC), which further improves the separation degree and analysis speed on the basis of HPLC, and mass spectrometry MS ionizes atoms or molecules according to the mass-to-charge ratio of atoms or molecules, thereby realizing the separation and detection of compounds, and has exclusive identification capabilities. MS has the characteristics of high selectivity and high sensitivity, which effectively makes up for the defects of insufficient selectivity and sensitivity of HPLC, avoids the problem that HPLC cannot effectively separate multiple compounds with similar structures and polarities due to co-elution, and lays a foundation for the subsequent simultaneous detection of four cytokinins with similar structures in litchi.
Description
Technical Field
The invention relates to the technical field of phytohormone detection, in particular to a method for simultaneously detecting four cytokinins in litchi by using UPLC-MS/MS.
Background
Currently, enzyme-linked immunosorbent assay (ELISA) and High Performance Liquid Chromatography (HPLC) are mostly adopted for measuring cytokinin on litchi.
However, the existing detection method has the defects that ELISA method utilizes the specific reaction of antigen and antibody to build the relationship between the to-be-detected object and enzyme, and then the enzyme and the substrate generate color reaction to quantitatively detect. However, since a class of hormones is often composed of a plurality of substances with similar structures, when antigens and antibodies are combined, there is a situation of misidentification, and the hormones are also easily affected by external environment to cause errors, and the specificity and sensitivity are required to be improved.
HPLC methods perform separation and purification in a chromatographic column by utilizing the different interactions of sample components with two different phases (stationary and mobile). Since HPLC is fast and depends on the different polarities of the compounds, two compounds of similar structure and polarity can leave the chromatograph at the same or nearly the same time, resulting in co-flow which results in an inability to determine exactly at what point the mixture is flowing out. HPLC is a powerful separation but is not selective and sensitive enough.
Disclosure of Invention
The invention aims to avoid the defects in the prior art and provide a method for simultaneously detecting four cytokinins in litchi by UPLC-MS/MS, the method can accurately and simultaneously detect four cytokinins in litchi, and has the advantages of easiness in operation, short detection period, good reproducibility, high sensitivity and low cost.
In order to achieve the above purpose, the present invention provides the following technical solutions:
provides a method for simultaneously detecting four cytokinins in litchi by UPLC-MS/MS, which comprises the following steps,
Step one, extraction and purification, comprising:
S11, obtaining litchi sample tissues, and grinding the litchi sample tissues under the condition of liquid nitrogen to obtain sample powder;
s12, placing the sample powder into a centrifuge tube, adding acetonitrile solution, leaching for 6-10 hours at 3-4 ℃, shaking at regular time, centrifuging the centrifuge tube at 3-4 ℃, collecting supernatant and residues,
Wherein 5-11 mL of acetonitrile solution is added to each 1g of sample powder, and the concentration of the acetonitrile solution is 60-85%;
s13, repeating the step S12 on the residues, and recovering the supernatant obtained in the residues and combining the supernatant obtained in the step S12 to obtain a sample crude extract;
S14, sequentially activating and balancing the PCX column by using methanol and 2% formic acid aqueous solution, eluting by using an eluent, collecting the eluent,
Wherein the leaching solution consists of a methanol solution and formic acid, the content of the formic acid is 0.05-0.2%,
The eluent consists of a methanol solution and ammonia water, wherein the content of the ammonia water is 1-3%, and 15-21 mL of the eluent is added into each 1g of sample powder;
s15, blowing and concentrating the eluting liquid nitrogen, and then adding a methanol aqueous solution for re-dissolving to obtain a re-solution, wherein the concentration of the methanol aqueous solution is 10% -20%;
step two, UPLC-MS/MS detection analysis, which comprises:
S21, carrying out UPLC-MS/MS detection analysis on the complex solution,
The liquid phase condition is that the mobile phase B is methanol, the mobile phase A is 5mmol/L ammonium formate aqueous solution, the elution is carried out by a gradient elution mode, the elution program is shown in the table 1,
TABLE 1 gradient elution procedure
The mass spectrum conditions are that an electrospray ion source is adopted, the detection mode is a multi-reaction detection mode, the temperature of the ion source is 150 ℃, the capillary voltage is 0.35kV, the desolvation gas temperature is 500 ℃, the atomizing gas flow rate is 1000L/Hr, the taper hole gas flow is 50L/Hr, the collision argon is 0.17L/h, the relevant mass spectrum parameters of four cytokinins are shown in the table 2,
Table 2 relevant mass spectral parameters for four cytokinins
In some embodiments, in S12, shake is performed 1 to 3 times.
In some embodiments, in S12, the centrifugal speed is 9000-11000 r/min and the centrifugal time is 7-15 min.
In some embodiments, in S21, the complex solution is vortexed and filtered through a 0.22um organic filter before performing UPLC-MS/MS detection analysis.
The method for simultaneously detecting four cytokinins in litchi by using UPLC-MS/MS has the beneficial effects that:
The method for simultaneously detecting four cytokinins in litchi by using liquid-phase secondary mass spectrometry (UPLC-MS/MS) adopts ultra-high performance liquid chromatography (UPLC), UPLC further improves the separation degree and the analysis speed on the basis of HPLC, and mass spectrometry MS ionizes atoms or molecules according to the mass-to-charge ratio of the atoms or molecules, so that separation and detection of the compounds are realized, the method has the characteristics of special identification capability, high selectivity and high sensitivity, the defect of insufficient HPLC selectivity and sensitivity is effectively overcome, the problem that various compounds with similar structures and polarities cannot be effectively separated due to co-outflow of HPLC is avoided, and a basis is laid for subsequently simultaneously detecting the four cytokinins with similar structures in litchi. The UPLC-MS/MS integrates the advantages of HPLC and MS, has the characteristics of high separability and high speed of UPLC, and has the characteristics of high selectivity and high sensitivity of MS, the invention has the advantages of easy operation of pretreatment, short detection period, good reproducibility, high sensitivity, low cost and the like, and the final result is reliable, thereby being applicable to rapid screening and quantitative determination of cytokinin in litchi and being applied to practical work.
Drawings
FIG. 1 is a flow chart of a method for simultaneously detecting four cytokinins in litchi by UPLC-MS/MS according to an embodiment of the invention.
Detailed Description
Preferred embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. While the preferred embodiments of the present invention are shown in the drawings, it should be understood that the present invention may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.
Examples
Referring to fig. 1, the present example discloses a method for simultaneously detecting four cytokinins in litchi by using UPLC-MS/MS, comprising the following steps,
Step one, extraction and purification, comprising:
S11, obtaining litchi sample tissues, and grinding the litchi sample tissues under the condition of liquid nitrogen to obtain sample powder;
grinding is carried out under the protection of liquid nitrogen, so that the sample is prevented from being oxidized.
S12, placing the sample powder into a centrifuge tube, adding acetonitrile solution (ACN), leaching for 6-10 hours at 3-4 ℃, shaking at regular time, centrifuging the centrifuge tube at 3-4 ℃, collecting supernatant and residues,
Wherein 5-11 mL of acetonitrile solution is added to each 1g of sample powder, and the concentration of the acetonitrile solution is 60-85%;
Cytokinin in the sample powder was collected into acetonitrile solution by leaching with acetonitrile solution. The shaking mode can effectively improve the leaching effect.
S13, repeating the step S12 on the residues, and recovering the supernatant obtained in the residues and combining the supernatant obtained in the step S12 to obtain a sample crude extract;
The residue is repeatedly leached to achieve the maximum possible recovery of cytokinins in the residue.
S14, sequentially activating and balancing the PCX column by using Methanol (MEOH) and a 2% formic acid aqueous solution (Foxmic acidwater), eluting by using an eluent, collecting the eluent,
Wherein the eluent consists of a Methanol (MEOH) solution and formic acid (Foxmic acid), the content of the formic acid is 0.05-0.2%,
The eluent consists of a methanol solution and ammonia water (ammonia), wherein the content of the ammonia water is 1-3%, and 15-21 mL of the eluent is added into each 1g of sample powder;
purification is performed by the steps of activation, equilibration, rinsing and elution.
S15, blowing and concentrating the eluting liquid nitrogen, and then adding a methanol aqueous solution for re-dissolving to obtain a re-solution, wherein the concentration of the methanol aqueous solution is 10-20%;
Purifying, concentrating, and re-dissolving to a specific volume by re-dissolving.
Step two, UPLC-MS/MS detection analysis, which comprises:
s22, carrying out UPLC-MS/MS detection analysis on the complex solution,
The liquid phase condition is that the mobile phase B is methanol, the mobile phase A is 5mmol/L ammonium formate aqueous solution, the elution is carried out by a gradient elution mode, the elution program is shown in the table 1,
TABLE 1 gradient elution procedure
The mass spectrum conditions are that an electrospray ion source is adopted, the detection mode is a multi-reaction detection mode, the temperature of the ion source is 150 ℃, the capillary voltage is 0.35kV, the desolvation gas temperature is 500 ℃, the atomizing gas flow rate is 1000L/Hr, the taper hole gas flow is 50L/Hr, the collision argon is 0.17L/h, the relevant mass spectrum parameters of four cytokinins are shown in the table 2,
Table 2 relevant mass spectral parameters for four cytokinins
The components of the sample after liquid chromatography separation sequentially enter a mass spectrum detector, and are ionized in an ion source to generate ions with certain charges and different mass numbers. The motion behaviors of different ions in the electromagnetic field are different, and the ions are separated according to different mass-to-charge ratios (m/z) by adopting a mass analyzer, so that mass spectrograms which are arranged according to the sequence of the mass-to-charge ratios are obtained. And (5) obtaining qualitative and quantitative results of the sample through analysis and treatment of a mass spectrogram.
In this embodiment, in S12, shake is performed 1 to 3 times.
In the embodiment, in S12, the centrifugal speed is 9000-11000 r/min, and the centrifugal time is 7-15 min.
In this embodiment, in S21, the complex solution is vortexed and uniformly mixed, and then passed through an organic filter membrane of 0.22um, and then subjected to UPLC-MS/MS detection analysis.
Impurity removal is achieved through the organic filter membrane, and detection accuracy is improved.
Experimental example
In order to verify the effect of the invention in detecting four cytokinins of litchi, the following experiment is carried out, and the steps comprise:
extraction and purification, namely 0.25g of litchi sample ground by liquid nitrogen is taken in a 15mL plastic centrifuge tube, 2.5mL of 80% acetonitrile solution is added, and leaching is carried out for 8 hours at 4 ℃ and shaking is carried out for 2 times.
Taking out, centrifuging at 4deg.C at 10000r/min for 10min, collecting supernatant, and collecting residue.
Adding 2.5mL of 80% acetonitrile into the residue, oscillating for 10min, centrifuging at 4 ℃ at 10000r/min for 10min, and combining the two extracts to obtain a crude extract of the sample.
Sequentially activating and balancing with methanol and 2% formic acid aqueous solution, eluting with methanol solution (containing 0.1% formic acid), eluting with 2.5ml2% ammonia water-methanol solution twice, collecting eluate in test tube, blowing nitrogen to near dryness, adding 0.25ml 5% methanol aqueous solution, re-dissolving, vortex mixing, and passing through 0.22um organic filter membrane to be tested.
Liquid phase and Mass Spectrometry conditions in liquid phase conditions mobile phase B was methanol and mobile phase A was 5mmol/L aqueous ammonium formate, elution gradients were as shown in Table 1 above.
The mass spectrum condition is electrospray ion source (ESI+), and the detection mode is a multi-reaction detection (MRM) mode. The ion source temperature is 150 ℃, the capillary voltage is 0.35kV, the desolvation gas temperature is 500 ℃, the atomizing gas flow rate is 1000L/Hr, the taper hole gas flow is 50L/Hr, and the collision gas (argon) is 0.17L/h. Other mass spectral parameters are shown in table 2 above.
Analysis of results:
As can be seen from tables 3 and 4, the detection limit (3 times signal to noise ratio) and the quantification limit (10 times signal to noise ratio) of the four cytokinins (TZR, IPR, DHZ, IP) are respectively lower than 9.35 and 31.15pg/g, and have good linear relation within the concentration range of 0.05-50 ng/mL, and the linear correlation coefficient (r 2) is higher than 0.999. The average recovery rate of the four cytokinins is 80.0% -108.2% and the standard deviation is 0.8% -15.5% under the standard adding level of three kinds of high, medium and low concentrations (0.4,2 and 20 ng/mL).
Therefore, the invention can efficiently and simultaneously detect four cytokinins in litchi.
Table 3 labeling recovery rates and relative standard deviation% of four cytokinins in litchi rind
TABLE 4 Linear equation, linear Range, correlation coefficient and detection Limit for four cytokinins
The analysis of the results effectively demonstrates the feasibility of the method for simultaneously detecting four cytokinins in litchi by using UPLC-MS/MS.
According to the technical scheme, four cytokinins of trans-zeatin nucleoside (TZR), dihydrozeatin (DHZ), isopentenyl adenine (IP) and isopentenyl adenine nucleoside (IPR) in litchi are measured simultaneously by using a liquid-phase secondary mass spectrometry (UPLC-MS/MS) method. 0.25g litchi sample was leached with acetonitrile-water (80:20, v/v) solution for 8h, purified by Bond Elut Plexa PCX solid phase extraction column (PCX column), eluted with 2.5% aqueous ammonia methanol and detected by 0.22um organic filter. And selecting XSelect HSS T chromatographic columns, carrying out gradient elution for 7min by taking methanol and 5mmol/L ammonium formate aqueous solution as mobile phases (table 1), adopting electrospray positive ion (ESI+) mode ionization, and selecting a reaction monitoring (MRM) mode to quantify cytokinin, so as to realize simultaneous detection and analysis of four cytokinins in litchi with similar structures.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
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| CN103822983A (en) * | 2014-03-03 | 2014-05-28 | 中国科学院武汉植物园 | Efficient turfgrass endogenous hormone separation and determination method |
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