CN109324139A - Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf - Google Patents
Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf Download PDFInfo
- Publication number
- CN109324139A CN109324139A CN201811538574.9A CN201811538574A CN109324139A CN 109324139 A CN109324139 A CN 109324139A CN 201811538574 A CN201811538574 A CN 201811538574A CN 109324139 A CN109324139 A CN 109324139A
- Authority
- CN
- China
- Prior art keywords
- liquid
- ribosylzeatin
- measuring method
- extraction
- solution
- 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.)
- Pending
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N30/14—Preparation by elimination of some components
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N2030/062—Preparation extracting sample from raw material
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N30/14—Preparation by elimination of some components
- G01N2030/146—Preparation by elimination of some components using membranes
Abstract
Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf, the following steps are included: with methanol: water: formic acid solution (80:19.9:0.1, V/V/V) it is used as leaching liquor, tobacco sample is after low temperature extracts, the extraction of ethyl acetate low temperature ultrasonic is added in leaching liquor, after extract liquor low-temperature centrifugation, take upper organic phase clear liquid, cryogenic nitrogen blows close dry, through organic membrane filtration after methanol redissolution, using the ribosylzeatin in Liquid Chromatography-tandem Mass method measurement tobacco leaf, inner mark method ration.The invention has the characteristics that avoiding the influence of analytic process mesostroma effect using Isotope internal standard dilution standard measure；This method pre-treatment is simple, can quickly, accurately detect the content of ribosylzeatin in tobacco leaf, and repeatability and the rate of recovery are good, the analysis suitable for batch samples.
Description
Technical field
The invention belongs to physical and chemical inspection technical fields, and in particular to the measuring method of ribosylzeatin content in tobacco leaf.
Background technique
Ribosylzeatin is the important form that the basic element of cell division transports in xylem, and the basic element of cell division is aza ring
Alkaline small, major function be promote tobacco leaf cells division and expansion, some researches show that the basic elements of cell division to pass through
The synthesis of the accumulation and protein and nucleic acid that promote substance prevents cigarette senescence of leaves, and the basic element of cell division is wide in tobacco plant body
General distribution.The current detection method in relation to the tobacco endogenous plant basic element of cell division mainly has immunoassay and chromatography.Its
Middle immunoassay has the advantage of lower detection limit, but need to solve the problems, such as the cross reaction of the antibody of preparation；Chromatography at present
With the most extensively, although the analysis methods such as gas chromatography-mass spectrum, high performance liquid chromatography have been able to plant tissue or cell
Plant hormone in extracting solution carries out quantitative analysis, but for detecting while plant hormone in a small amount of even trace plant sample
There is also difficulties.Main cause, first is that the content of ribosylzeatin hormone is extremely low, it is so low usually all in the ng/g order of magnitude
Its detection method of content requirement and instrument have the sensitivity of superelevation to plant hormone；Second is that ribosylzeatin hormone it is unstable,
It is more sensitive to temperature and media environment；Third is that tobacco plant system matrix is sufficiently complex, directly detection will receive a large amount of matrix
The influence of chaff interferent.Therefore there is very high requirement to the selectivity and purification capacity of sample treatment, while requiring quantitative point
Analysis method and instrument have the sensitivity of superelevation to target plant hormone.But it is in recent years, quick with mass spectrometric hyphenated technique with chromatography
Development, in tandem mass spectrum (MS/MS) parent ion and the one-to-one multiple reaction monitoring pattern of daughter ion can effectively remove because
False positive caused by complex matrices interference, relative to single-stage mass spectrometry method and high performance liquid chromatography, LC-MS/MS method is a kind of
Sensitivity is higher, and selective more preferable, the stronger method of specificity has well in terms of analysis speed, sensitivity, selectivity
Advantage.Therefore, by carrying out pre-treatment appropriate to this complex sample system of tobacco, to the selective enumeration method energy of plant hormone
Power is greatly improved, and can be realized accurate qualitative.
The liquid-liquid extraction of ribosylzeatin-not disclosed report of liquid chromatography-tandem mass spectrometry analysis method in tobacco leaf, accurately
Its content is measured, can be the physiological function and metabolic pathway for being best understood from the main Endogenous hormones of tobacco, be tobacco planting
Technical support is provided, base yield of tobacco and quality are improved.
Summary of the invention
The present invention compensates for the deficiency of existing analytical technology, provides ribosylzeatin liquid-liquid extraction-liquid phase in a kind of tobacco leaf
Chromatography-tandem mass spectrum measuring method.
The technical solution of the present invention is to provide ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometries in a kind of tobacco leaf
Measuring method, comprising the following steps:
(1) methanol, water and formic acid, compounding methanol: water: formic acid solution (80:19.9:0.1, V/ the preparation of extracting solution: are used
V/V) it is used as extracting solution.
(2) internal standard stock solution is prepared: preparing deuterated ribosylzeatin (d2-ZR) internal standard stock solution with methanol, concentration is
1000ng/mL。
(3) standard reserving solution is prepared: preparing ribosylzeatin (ZR) standard reserving solution with methanol, concentration is 100 μ g/mL.
(4) sample pre-treatments and analysis: fresh tobacco leaves sample is stored in spare in -50 DEG C of refrigerators after pulverized under liquid nitrogen.
The fresh tobacco leaves sample that takes that treated in 2mL centrifuge tube, be added internal standard stock solution (1000ng/mL) and extracting solution (methanol:
Water: formic acid solution=80:19.9:0.1, V/V/V), low temperature extraction is rear that the extraction of extractant low temperature ultrasonic is added, and extract liquor passes through
After centrifugation, about 900 μ L of supernatant liquor is pipetted, nitrogen, which blows to be concentrated into, closely to be done, it is redissolved with methanol, after redissolution liquid crosses organic film, sample examination
Liquid carries out LC-MS/MS measurement.
(5) Specification Curve of Increasing: the working stamndard that blank sample liquid dilution standard stock solution is made into series of concentrations is molten
Liquid makes standard curve.The series of concentrations of Working Standard Solution configuration is 1,5,10,20,50,100,200ng/m L, obtains 7 grades
Working Standard Solution；ZR and d2-ZR uses positive ion mode, LC-MS/MS analysis is carried out to above-mentioned series standard solution, with mesh
It marks object and linear regression analysis is carried out to the corresponding concentration proportion of the two with interior target peak area ratio, acquire the mark of ribosylzeatin
Directrix curve.
(6) inner mark method ration data processing and detection: is carried out by the ratio between target peak area and internal standard peak area.
Further, in step (4), the tobacco sample amount that weighs that treated and internal standard stock solution and extracting solution is added
Total volume ratio is 1:10 (mg/ μ L).
Further, in step (4), weighed tobacco sample amount is 50mg.
Further, in step (4), the standard substance in internal standard stock solution is deuterated ribosylzeatin (d2-ZR).
Further, in step (4), the additional amount of internal standard stock solution and extract liquor is respectively 50 μ L and 500 μ L.
Further, in step (4), extraction time 12h, extraction temperature is 4 DEG C.
Further, in step, extractant is ethyl acetate, usage amount 1mL, extraction time 15min, extraction temperature
Degree is 4 DEG C.
Further, in step (4), centrifugally operated condition are as follows: centrifugal rotational speed 10000r/min, centrifugation time are
5min, centrifuging temperature are 4 DEG C.
Further, in step (4), organic filter membrane is 0.22 μm of teflon membrane filter.
Further, in step (4), matched using determining without ribosylzeatin blank sample liquid dilution standard stock solution
At series of concentrations.
Further, in step (4) or (5), when LC-MS/MS is measured, high-efficient liquid phase chromatogram condition are as follows:
Chromatographic column: Waters Symmetry C18, column parameter are 150mm × 3.9mm × 5 μm；
Pre-column: Waters Symmetry C18, column parameter 10mm × 3.9mm × 5 μm；
Sample volume: 20 μ L；Column temperature: 25 DEG C；Flow velocity is 0.5mL/min；
Mobile phase A is 0.1% formic acid；Mobile phase B is methanol: formic acid=99.9:0.1 (V/V)
Gradient elution program are as follows:
Further, in step (4) or (5), Mass Spectrometry Conditions are as follows: Mass Spectrometer Method uses ESI ion source, sweeps in cation
It retouches under mode, MRM operating mode is selected to carry out second mass analysis, the main Mass Spectrometry Conditions after optimization are as follows: under positive ion mode,
Electron spray voltage 5500V；Atomization gas 50psi, auxiliary heating gas 50psi, gas curtain gas 15psi, 600 DEG C of drying temperature；Object
And interior target characteristic ion and parameter are respectively as follows:
The beneficial effects of the present invention are:
The instrument and equipment that the present invention uses is laboratory conventional analysis equipment, and inventive method is easy to spread, detection at
This is low；
The present invention avoids complex object system Quantitative Separation, the difficulty of purifying using Isotope internal standard dilution standard measure,
Overcome the influence of matrix effect；
Present invention firstly provides using zeatin in liquid-liquid extraction-liquid chromatography-tandem mass instrument measurement tobacco leaf
The analysis method of nucleosides, method analysis speed is fast, and repeatability and the rate of recovery are good, the analysis suitable for batch samples.
Detailed description of the invention
Fig. 1 show the extraction ion flow graph of ribosylzeatin (ZR).
Specific embodiment
To facilitate the understanding of the present invention, the present invention further illustrates in conjunction with specific embodiments:
Instrument and reagent, material: 4500 LC-MS/MS of American AB SCIEX；U.S. Thermo U3000 HPLC；
Waters Symmetry C18 chromatographic column (150mm × 3.9mm, 5 μm of partial size), 0.22 μm of micropore filtering film, low temperature ultrasonic instrument；
Ultrapure water (18M Ω), ethyl acetate, methanol, formic acid (chromatographically pure, lark prestige chemical reagents corporation), ribosylzeatin and its interior
Mark standard substance (Town in Shanghai spectrum experiment Science and Technology Co., Ltd.).
The preparation of extracting solution: methanol, water and formic acid, compounding methanol: water: formic acid solution (80:19.9:0.1, V/V/ are used
V) it is used as extracting solution.
It prepares internal standard stock solution: preparing deuterated ribosylzeatin (d2-ZR) internal standard stock solution with methanol, concentration is
1000ng/mL。
Preparing standard solution: ribosylzeatin (ZR) standard reserving solution is prepared with methanol, concentration is 100 μ g/mL.
Sample pre-treatments: fresh tobacco leaves sample is stored in spare in -50 DEG C of refrigerators after pulverized under liquid nitrogen.It takes at about 50mg
50 μ L internal standard working solutions (1000ng/mL) and the methanol of 500 μ L is added in 2mL centrifuge tube in fresh tobacco leaves sample after reason:
Water: formic acid solution (80:19.9:0.1, V/V/V) extracts 12 hours in 4 DEG C, and the rear 1000 μ L ethyl acetate that are added are in 4 DEG C of ultrasound extractions
15min is taken, extract liquor is centrifuged 5min in 4 DEG C with 10000r/min, pipettes about 900 μ L of supernatant liquor, and nitrogen, which is blown, is concentrated into close dry, use
100 μ L methanol redissolve, and cross 0.22 μm of organic filter membrane, and sample test solution waits for that LC-MS/MS is measured.
High-efficient liquid phase chromatogram condition:
Chromatographic column: Waters Symmetry C18, column parameter are 150mm × 3.9mm × 5 μm；
Pre-column: Waters Symmetry C18, column parameter 10mm × 3.9mm × 5 μm；
Sample volume: 20 μ L；Column temperature: 25 DEG C；Flow velocity is 0.5mL/min；
Mobile phase A is 0.1% formic acid；Mobile phase B is methanol: formic acid=99.9:0.1 (V/V)；
Gradient elution program is as shown in table 1:
1 Gradient Elution condition of table
Mass Spectrometry Conditions are as follows: Mass Spectrometer Method use ESI ion source, under cation scanning mode, select MRM operating mode into
Row second mass analysis, the main Mass Spectrometry Conditions after optimization are as follows: under positive ion mode, electron spray voltage 5500V；Atomization gas
50psi, auxiliary heating gas 50psi, gas curtain gas 15psi, 600 DEG C of drying temperature；Object and interior target characteristic ion and parameter
It is shown in Table 2:
2 ribosylzeatin of table and interior target characteristic ion and parameter
The drafting of standard curve:
It is molten that series of concentrations working stamndard is made into the determining blank sample liquid dilution standard stock solution without ribosylzeatin
Liquid makes standard curve.The series of concentrations of Working Standard Solution configuration is 1,5,10,20,50,100,200ng/m L, obtains 7 grades
Working Standard Solution；ZR and d2-ZR uses positive ion mode, LC-MS/MS analysis is carried out to above-mentioned series standard solution, with mesh
It marks object and linear regression analysis is carried out to the corresponding concentration proportion of the two with interior target peak area ratio, acquire the mark of ribosylzeatin
Directrix curve；The equation of linear regression of ribosylzeatin, related coefficient, detection limit and quantitative limit are as shown in table 3:
Equation of linear regression, related coefficient, detection limit and the quantitative limit of 3 ribosylzeatin of table
| Compound | Equation of linear regression | R2 | Detection limit (ng/g) | Quantitative limit (ng/g) |
| ZR | Y=13.55x+0.7056 | 0.9994 | 0.104 | 0.347 |
Precision: the precision in order to investigate method in a few days repeats to test, measurement result table with tobacco sample progress 6 times
The relative standard deviation of bright ribosylzeatin in a few days measurement result shows the in a few days accurate of method between 4.28%~6.70%
Degree is good.Meanwhile sample has been carried out 6 days repeating to test in the daytime, measurement result shows ribosylzeatin measurement result in the daytime
Relative standard deviation is between 5.65%~9.17%, the results showed that the repeatability in the daytime of this method is good.
The rate of recovery: the rate of recovery has been carried out on high, medium and low three contents levels to tobacco sample using sample mark-on method
Measurement, the results showed that, for the result of the average recovery rate of ribosylzeatin between 81.6%-109.4%, the rate of recovery is higher, full
Foot analysis requires.
The measurement of actual sample: the ribosylzeatin (table 4) of 4 tobacco leaves, sample are determined using the method that the present invention establishes
The content range of ribosylzeatin is 4.0-10.7ng/kg in product.
The testing result (ng/g) of ribosylzeatin in 4 tobacco leaf of table
| Compound | 1# | 2# | 3# | 4# |
| ZR | 6.3 | 4.0 | 9.1 | 10.7 |
Claims (12)
1. ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf, it is characterised in that:
Method includes the following steps:
(1) methanol, water and formic acid, compounding methanol: water: formic acid solution (80:19.9:0.1, V/V/V) preparation of extracting solution: are used
As extracting solution；
(2) internal standard stock solution is prepared: preparing deuterated ribosylzeatin (d2-ZR) internal standard stock solution with methanol, concentration is
1000ng/mL；
(3) standard reserving solution is prepared: preparing ribosylzeatin (ZR) standard reserving solution with methanol, concentration is 100 μ g/mL；
(4) sample pre-treatments and analysis: fresh tobacco leaves sample is stored in spare in -50 DEG C of refrigerators after pulverized under liquid nitrogen；It weighs
Treated fresh tobacco leaves sample in 2mL centrifuge tube, be added internal standard stock solution (1000ng/mL) and extracting solution (methanol: water:
Formic acid=80:19.9:0.1, V/V/V), low temperature extraction is rear that the extraction of extractant low temperature ultrasonic is added, extract liquor after centrifugation,
About 900 μ L of supernatant liquor is pipetted, nitrogen, which blows to be concentrated into, closely to be done, and is redissolved with methanol, after redissolution liquid crosses organic film, sample test solution is carried out
LC-MS/MS measurement；
(5) drafting of standard curve: with blank sample liquid dilution standard stock solution be made into series of concentrations be 1,5,10,20,50,
100, the 7 level work standard solution of 200ng/mL；ZR and d2-ZR uses positive ion mode, carries out to above-mentioned series standard solution
LC-MS/MS analysis carries out linear regression analysis to the corresponding concentration proportion of the two with interior target peak area ratio with object,
Acquire the standard curve of ribosylzeatin；
(6) inner mark method ration data processing and detection: is carried out by the ratio between target peak area and internal standard peak area.
2. the measuring method of ribosylzeatin in tobacco leaf according to claim 1, it is characterised in that: in step (4), institute
Stating weighed tobacco sample amount and the volume ratio of extracting solution is added is 1:10 (mg/ μ L).
3. measuring method according to claim 1, it is characterised in that: the reference substance in step (4), in internal standard stock solution
Matter is deuterated ribosylzeatin (d2-ZR).
4. measuring method according to claim 1, it is characterised in that: in step (4), internal standard stock solution and extracting solution
Additional amount is respectively 50 μ L and 500 μ L.
5. measuring method according to claim 1, it is characterised in that: in step (4), the weighed tobacco sample amount
For 50mg.
6. measuring method according to claim 1, it is characterised in that: in step (4), when the extraction of low temperature extraction
Between be 12h, extraction temperature be 4 DEG C.
7. measuring method according to claim 1, it is characterised in that: in step (4), extractant is ethyl acetate, is made
Dosage is 1mL, and extraction time 15min, extraction temperature is 4 DEG C.
8. measuring method according to claim 1, it is characterised in that: in step (4), the centrifugally operated condition are as follows:
Centrifugal rotational speed is 10000r/min, and centrifugation time 5min, centrifuging temperature is 4 DEG C.
9. measuring method according to claim 1, it is characterised in that: in step (4), organic filter membrane is 0.22 μm
Teflon membrane filter.
10. measuring method according to claim 1, it is characterised in that: in step (5), the blank sample liquid is free of
There is ribosylzeatin.
11. measuring method according to claim 1, it is characterised in that: in step (4) or (5), the LC-MS/MS
When measurement, high-efficient liquid phase chromatogram condition are as follows:
Chromatographic column: Waters Symmetry C18, column parameter are 150mm × 3.9mm × 5 μm；
Pre-column: Waters Symmetry C18, column parameter 10mm × 3.9mm × 5 μm；
Sample volume: 20 μ L；Column temperature: 25 DEG C；Flow velocity is 0.5mL/min；
Mobile phase A is 0.1% formic acid；Mobile phase B is methanol: formic acid=99.9:0.1 (V/V)；
Gradient elution program are as follows:
12. measuring method according to claim 1, it is characterised in that: in step (4) or (5), the LC-MS/MS is surveyed
Periodically, Mass Spectrometry Conditions are as follows: Mass Spectrometer Method uses ESI ion source, under cation scanning mode, MRM operating mode is selected to carry out
Second mass analysis, the main Mass Spectrometry Conditions after optimization are as follows: under positive ion mode, electron spray voltage 5500V；Atomization gas 50psi,
Auxiliary heating gas 50psi, gas curtain gas 15psi, 600 DEG C of drying temperature；Object and interior target characteristic ion and parameter are respectively as follows:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811538574.9A CN109324139A (en) | 2018-12-14 | 2018-12-14 | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811538574.9A CN109324139A (en) | 2018-12-14 | 2018-12-14 | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109324139A true CN109324139A (en) | 2019-02-12 |
Family
ID=65256181
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811538574.9A Pending CN109324139A (en) | 2018-12-14 | 2018-12-14 | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109324139A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112684034A (en) * | 2020-12-10 | 2021-04-20 | 中国烟草总公司郑州烟草研究院 | Non-target analysis method for detecting tobacco and tobacco products based on HPLC-QTOF MS technology |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6927062B2 (en) * | 2002-11-25 | 2005-08-09 | Agdia, Inc. | Controls and standards for assays and method for manufacture thereof |
| CN103048404A (en) * | 2012-12-19 | 2013-04-17 | 武汉大学 | Quantitative detection method for endogenous cytokinins in plant sample |
| CN103353490A (en) * | 2013-06-24 | 2013-10-16 | 浙江大学 | Method for detecting pear endogenous hormone by using ultra-high performance liquid chromatography electrospray tandem mass spectrum |
| US20140147925A1 (en) * | 2011-04-15 | 2014-05-29 | Basf Plant Science Company Gmbh | Method for Profiling Phytohormone Levels in Plant Tissue |
| CN105486788A (en) * | 2015-12-31 | 2016-04-13 | 苏州科铭生物技术有限公司 | Kit for determining zeatin nucleoside content and method thereof |
| CN106501427A (en) * | 2016-10-26 | 2017-03-15 | 武汉绿剑可瑞信科技有限公司 | The pre-treating method and simultaneous quantitative determination of multiple endogenous plant hormones in a kind of plant sample |
| CN107703238A (en) * | 2017-10-27 | 2018-02-16 | 四川农业大学 | HPLC methods that are a kind of while determining 6 kinds of Endogenous Hormone Contents in Vitro in the red sage root |
| 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 |
-
2018
- 2018-12-14 CN CN201811538574.9A patent/CN109324139A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6927062B2 (en) * | 2002-11-25 | 2005-08-09 | Agdia, Inc. | Controls and standards for assays and method for manufacture thereof |
| US20140147925A1 (en) * | 2011-04-15 | 2014-05-29 | Basf Plant Science Company Gmbh | Method for Profiling Phytohormone Levels in Plant Tissue |
| CN103048404A (en) * | 2012-12-19 | 2013-04-17 | 武汉大学 | Quantitative detection method for endogenous cytokinins in plant sample |
| CN103353490A (en) * | 2013-06-24 | 2013-10-16 | 浙江大学 | Method for detecting pear endogenous hormone by using ultra-high performance liquid chromatography electrospray tandem mass spectrum |
| CN105486788A (en) * | 2015-12-31 | 2016-04-13 | 苏州科铭生物技术有限公司 | Kit for determining zeatin nucleoside content and method thereof |
| CN106501427A (en) * | 2016-10-26 | 2017-03-15 | 武汉绿剑可瑞信科技有限公司 | The pre-treating method and simultaneous quantitative determination of multiple endogenous plant hormones in a kind of plant sample |
| 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 |
| CN107703238A (en) * | 2017-10-27 | 2018-02-16 | 四川农业大学 | HPLC methods that are a kind of while determining 6 kinds of Endogenous Hormone Contents in Vitro in the red sage root |
Non-Patent Citations (4)
| Title |
|---|
| GROLAMYS CASTILLO 等: "Simultaneous quantification of phytohormones in fermentation extracts of Botryodiplodia theobromae by liquid chromatography-electrospray tandem mass spectrometry", 《WORLD J MICROBIOL BIOTECHNOL》 * |
| KLARA HOYEROVA 等: "Efficiency of different methods of extraction and purification of cytokinins", 《PHYTOCHEMISTRY》 * |
| 刘雪梅 等: "LC-MS同时测定大型海藻中9个植物激素", 《药物分析杂志》 * |
| 庞夫花: "日中性草莓花芽分化及生化物质变化研究", 《中国优秀硕士学位论文全文数据库 农业科技辑（月刊）》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112684034A (en) * | 2020-12-10 | 2021-04-20 | 中国烟草总公司郑州烟草研究院 | Non-target analysis method for detecting tobacco and tobacco products based on HPLC-QTOF MS technology |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106442758A (en) | Liquid mass spectrometry method for detecting various amino acids in human blood plasma in underivatized mode | |
| CN108918711A (en) | The detection method of polyphenol compound in a kind of tobacco leaf | |
| CN105675788B (en) | The method of progesterone and testosterone in high performance liquid chromatography tandem mass spectrum technology for detection saliva | |
| CN105891364A (en) | Method and kit for detecting melatonin in saliva with HPLC-MS/MS (high performance liquid chromatography-tandem mass spectrometry) technology | |
| CN105259287A (en) | Second-order mass spectrometric detection method for bisphenol substances in water environment | |
| CN106442836A (en) | Method for detecting contents of folic acid and sulfur-containing amino acid in plasma | |
| CN108872415A (en) | The analyzing detecting method of monohydroxy polycyclic aromatic hydrocarbon in a kind of urine | |
| CN106706826A (en) | Analysis method of plant hormones in milligram-grade plants | |
| CN109060983A (en) | A kind of method of liquid chromatography-tandem mass spectrometry detection metanephrine substance | |
| CN108362795A (en) | Content of homocysteine rapid detection method in dried blood spot | |
| CN113049719A (en) | Method and kit for detecting free testosterone | |
| CN112986433A (en) | Method for detecting steroid content in human serum sample | |
| CN108072712B (en) | Quantitative analysis method for blood concentration of new compound WSJ-557 in SD rat plasma | |
| CN112014509A (en) | Method for synchronously determining angiotensin I and aldosterone in sample | |
| CN102980968A (en) | Liquid chromatogram tandem mass spectrum measuring method for creatinine in urine | |
| CN111426776B (en) | Application of HQR as characteristic marker of schefflera octophylla honey | |
| CN112162054B (en) | Method for evaluating authenticity of sophora moorcroftiana honey | |
| CN110333308B (en) | Method for simultaneously determining NNAL and cotinine in urine with high sensitivity and accuracy | |
| CN109324139A (en) | Ribosylzeatin liquid-liquid extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf | |
| CN109324140A (en) | Ribosylzeatin Solid Phase Extraction-liquid chromatography-tandem mass spectrometry measuring method in a kind of tobacco leaf | |
| CN112526048B (en) | Method for rapidly detecting trace residues of hypertensive drugs in environmental sediment | |
| CN111060612B (en) | Method for determining concentration of prednisone, prednisone acetate and active metabolite prednisolone in blood plasma by liquid chromatography-mass spectrometry | |
| CN105699575A (en) | Method and kit for testing cortisol in saliva by efficient liquid chromatogram and tandem mass spectrometry combination technology | |
| CN113866315A (en) | Quantitative analysis method for detecting rat plasma YG-18 blood concentration by liquid chromatography-mass spectrometry technology | |
| CN112213417A (en) | Kit and method for detecting concentration of mycophenolic acid medicine in dried blood spots |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190212 |
|
| RJ01 | Rejection of invention patent application after publication |