CN110243959A - A kind of separation of endogenous fragrance component in Carnation flowers and identification method - Google Patents
A kind of separation of endogenous fragrance component in Carnation flowers and identification method Download PDFInfo
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- CN110243959A CN110243959A CN201910503484.4A CN201910503484A CN110243959A CN 110243959 A CN110243959 A CN 110243959A CN 201910503484 A CN201910503484 A CN 201910503484A CN 110243959 A CN110243959 A CN 110243959A
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- 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
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- 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
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- 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
- G01N2030/042—Standards
- G01N2030/045—Standards internal
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- 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
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- 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
The invention belongs to plant component detection technique fields, and in particular to a kind of separation of the endogenous fragrance component in Carnation flowers and identification method.The present invention provides to the endogenous fragrance component of the Carnations flowers such as Chinese China pink, carnation, beared pink and dianthus superbus separation and identification method.The present invention separates and collects endogenous aroma substance in Carnation flowers including the use of organic solvent MTBE extracting process, Carnation flowers external source volatilization aroma substance is tested and analyzed using gas chromatography combined with mass spectrometry, and to fragrance component progress qualitative and quantitative analysis.In the qualitative analysis to the endogenous fragrance component of detection, unknown compound after retrieval with NIST MS Search2.3 spectrum library by matching, and with parameter SI > 800, the ingredient of RSI > 800 is as qualitative.The content of qualitative component is calculated by internal standard method.Separation and identification method of the invention has many advantages, such as that easy to operate, quick and applicability is wide.
Description
Technical field
The invention belongs to plant component technical field of analysis and detection, and in particular to the endogenous fragrance in a kind of Carnation flowers
The separation of ingredient and identification method.
Background technique
There are about 600 kinds of flowers under Carnation, common kind includes Chinese China pink, dianthus superbus, carnation (carnation) and palpus
Luxuriant China pink etc. is all that ornamental type is widely cultivated under Carnation.Growing and cultivation Carnation flowers can not only be seen for people
Reward can also extract essence, extract production cosmetics material requested, therefore will have to the analysis of fragrance component in Carnation flowers
Huge application, economic value.
Not yet have in domestic or even world wide for the analysis method of entire Carnation flowers fragrance of a flower substance at present
Whole system, it is only external to the aroma substance separating and extracting process of flowers has been reported in other categories under the Caryophyllaceae of part, therefore build
The aroma substance analysis system for standing complete Carnation flowers will have great theoretical and practice significance.
Summary of the invention
It is an object of the invention to overcome the deficiencies of existing technologies, for Carnation flower fragrance substance method of separating and assaying
Vacancy, the present invention at home and abroad provides the side that the fragrance component in a kind of pair of Carnation flowers is separated and identified for the first time
Method, to fill up the blank of Carnation flower fragrance elemental analysis.
It is described that technical scheme is as follows:
The present invention provides the separation of the fragrance component in a kind of pair of Carnation flowers and identification methods, including following step
It is rapid:
(1) endogenous aroma substance in Carnation flowers is separated and collected using organic solvent MTBE extracting process, specifically:
Take it is fresh, in liquid nitrogen be ground to powdery petal samples 0.10g in centrifuge tube, 500 μ L ultrapure waters are added, oscillation mixes；To
Be added in centrifuge tube 500 μ L MTBE (containing 5 × 10-3 μ g isobutylbenzene as internal standard), oscillation mix after, be added 0.2-0.3g without
Aqueous sodium persulfate removes moisture removal, and vibrates in vortex instrument and mix 2min；At room temperature, 2min is centrifuged with 12000rmp；With disposable
Syringe takes upper organic phase, is collected in sample injection bottle after excessively organic filter membrane, to be measured；
(2) the above-mentioned endogenous perfume (or spice) of gained Carnation flowers is tested and analyzed using the gas chromatography combined with mass spectrometry of fluid sample sample introduction
Gas substance, and qualitative and quantitative analysis is carried out to fragrance component therein；
A) GC conditions: using DB-5MS quartz capillary column, and the DB-5MS quartz capillary column parameter is 30m
× 0.250mm × 0.25 μm, carrier gas are helium, flow 1.0mL/min, using not shunt mode sample introduction, injector temperature
It is 250 DEG C, temperature program are as follows: initial temperature is 35 DEG C, keeps 4min, rises to 250 DEG C with 5 DEG C/min, keeps 15min；
B) Mass Spectrometry Conditions: electron impact ion source, electron energy 70eV, ion source temperature are 280 DEG C, ion transfer line
Temperature is 260 DEG C, using full scan mode, scanning range m/z:30-350；
(3) during the fragrance component of gas chromatography combined with mass spectrometry detection carries out qualitative analysis, unknown compound is through examining
Rope matches with NIST MS Search2.3 spectrum library simultaneously, and with parameter RSI > 800, the ingredient of SI > 800 is as qualitative parameter；
(4) in a dosing process, compounds content is calculated by internal standard method；I.e. with the peak area of various compounds
Divided by the peak area of internal standard substance isobutylbenzene multiplied by the content (μ g) of internal standard substance divided by sample quality (g)；
Wherein: the organic solvent MTBE, that is, methyl tertiary butyl ether(MTBE) is bought by SIGMA ALDRICH company
The analysis and detecting instrument is the gas chromatography-mass spectrometry suitable for fluid sample sample introduction.
The quantitative approach is inner mark method ration.
The positive effect of the present invention:
The present invention worldwide announces the technology for being suitable for Carnation flower fragrance species analysis for the first time, using solvent
Extraction combines the fragrance component of gas chromatography combined with mass spectrometry technology analysis Carnation flowers, and it is quantitative to form a set of fragrance component
Method for qualitative analysis.Identification method of the invention have it is easy to operate, quick, quantification and qualification is accurate, be suitable for China pink
All kinds of flowers of subordinate test and analyze.
Detailed description of the invention
Fig. 1: embodiment 1 uses organic solvent MTBE abstraction technique-fluid sample sample introduction gas chromatography combined with mass spectrometry skill
The chromatogram that art analyzes the endogenous fragrance component in Carnation flowers.
Specific embodiment
Embodiment 1
The present embodiment mainly uses organic solvent MTBE abstraction technique (Lavy M etc., 2002)-fluid sample sample introduction gas
Phase hydrolysis and condensation carries out qualitative, quantitative to the endogenous fragrance component in Carnation flowers, the specific steps are as follows:
1) endogenous fragrance component in Carnation flowers is extracted:
Take it is fresh, in liquid nitrogen be ground to powdery petal samples 0.10g in centrifuge tube, 500 μ L ultrapure waters, oscillation is added
It mixes；500 μ L are added into centrifuge tube and contain 5 × 10-3 μ g isobutylbenzene as internal standard), after oscillation mixes, 0.2-0.3g is added
Anhydrous sodium sulfate removes moisture removal, and vibrates in vortex instrument and mix 2min；At room temperature, 12000rmp is centrifuged 2min；With disposable
Syringe takes upper organic phase, is collected in sample injection bottle after excessively organic filter membrane, to be measured.
2) above-mentioned gained Carnation is tested and analyzed using the gas chromatography combined with mass spectrometry technology for being suitable for fluid sample sample introduction
Endogenous aroma substance in flowers, and qualitative and quantitative analysis is carried out to wherein fragrance component.
GC conditions:
Using DB-5MS quartz capillary column, the DB-5MS quartz capillary column parameter is 30m × 0.250mm × 0.25
μm, carrier gas is helium；Flow is 1.0mL/min；Using not shunt mode sample introduction, injector temperature is 250 DEG C；Temperature program
Are as follows: initial temperature is 35 DEG C, keeps 4min, rises to 250 DEG C with 5 DEG C/min, keeps 15min.
Mass Spectrometry Conditions:
Electron impact ion source, electron energy 70eV, ion source temperature are 280 DEG C, and ion transfer line temperature is 260
DEG C, using full scan mode, scanning range m/z:30-350.
3) qualitative and quantitative of substance is detected:
Unknown compound matches with NIST MS Search2.3 spectrum library simultaneously through retrieval, with parameter RSI > 800, SI >
800 ingredient is as qualitative.Table 1 is testing result of the present invention to endogenous fragrance component.
Wherein: the organic solvent MTBE, that is, methyl tertiary butyl ether(MTBE) is bought by SIGMA ALDRICH company.
The testing result of the endogenous aroma substance ingredient of 1 carnation of table
* content unit is 1 × 10-4μg/g
In the dosing process, compounds content is calculated by internal standard method；Within being removed with the peak area of various compounds
The peak area of mark substance isobutylbenzene is multiplied by the content (μ g) of internal standard substance divided by sample quality (g).
Leading reference:
Lavy M,Zuker A,Lewinsohn E,et al.Linalool and linalool oxide
production in transgenic carnation flowers expressing theClarkia
brewerilinalool synthase gene[J].Molecular Breeding,2002,9(2):103-111。
Claims (4)
1. separation and the identification method of a kind of endogenous fragrance component of Carnation flowers, feature the following steps are included:
(1) endogenous aroma substance in Carnation flowers is separated and collected using organic solvent MTBE extracting process, the specific steps are,
It takes petal samples 0.10g that is fresh, being ground in liquid nitrogen powdery Carnation flowers in centrifuge tube, 500 μ L ultrapure waters is added,
Oscillation mixes；It is added into centrifuge tube and contains 5 × 10-3μ g isobutylbenzene is as interior 500 μ L MTBE of target, after oscillation mixes, is added
0.2-0.3g anhydrous sodium sulfate removes moisture removal, is vibrated with vortex instrument and mixes 2min；At room temperature, 2min is centrifuged with 12000rmp；
Upper organic phase is taken with disposable syringe, is collected in sample injection bottle after excessively organic filter membrane, it is to be measured；
(2) endogenous fragrance object in gained Carnation flowers is tested and analyzed using the gas chromatography combined with mass spectrometry of fluid sample sample introduction
Matter, and qualitative and quantitative analysis, specific steps are carried out to wherein fragrance component are as follows:
GC conditions: use DB-5MS quartz capillary column, the DB-5MS quartz capillary column parameter be 30m ×
0.250mm × 0.25 μm, carrier gas are helium, and flow 1.0mL/min, using not shunt mode sample introduction, injector temperature is
250 DEG C, temperature program are as follows: initial temperature is 35 DEG C, keeps 4min, rises to 250 DEG C with 5 DEG C/min, keeps 15min；
Mass Spectrometry Conditions: electron impact ion source, electron energy 70eV, ion source temperature are 280 DEG C, and ion transfer line temperature is
260 DEG C, using full scan mode, scanning range m/z:30-350；
(3) during using the fragrance component qualitative analysis of gas chromatography-mass spectrum detection, unknown compound is through retrieving simultaneously
Match with NIST MS Search2.3 spectrum library, with parameter RSI > 800, the ingredient of SI > 800 is as qualitative；
(4) in a quantitative analysis process, compounds content is calculated by internal standard method, i.e., with the peak area of various compounds
Divided by the peak area of internal standard substance isobutylbenzene multiplied by the content (μ g) of internal standard substance divided by sample quality (g),
Wherein: the organic solvent MTBE, that is, methyl tertiary butyl ether(MTBE).
2. the isolation and identification method of the endogenous fragrance component of a kind of pair of Carnation flowers according to claim 1, feature exist
In: the analysis and detecting instrument is the gas chromatography-mass spectrometry of Liquid sample introduction.
3. a kind of Carnation flowers according to claim 1 are endogenous and the isolation and identification method of external source volatilization fragrance component,
It is characterized by: the quantitative approach is inner mark method ration.
4. the separation of the endogenous fragrance component in any one of -3 Carnation flowers and identification side according to claim 1
Method, which is characterized in that the Carnation flowers include Chinese China pink, carnation, beared pink and dianthus superbus.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112649539A (en) * | 2021-01-11 | 2021-04-13 | 湖南农业大学 | Detection method of in-vitro gas flower aroma substances |
| CN113759012A (en) * | 2020-08-27 | 2021-12-07 | 北京康仁堂药业有限公司 | Quality control method of dianthus superbus formula granules |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113759012A (en) * | 2020-08-27 | 2021-12-07 | 北京康仁堂药业有限公司 | Quality control method of dianthus superbus formula granules |
| CN113759012B (en) * | 2020-08-27 | 2023-03-21 | 北京康仁堂药业有限公司 | Quality control method of dianthus superbus formula granules |
| CN112649539A (en) * | 2021-01-11 | 2021-04-13 | 湖南农业大学 | Detection method of in-vitro gas flower aroma substances |
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Application publication date: 20190917 |