CN108663453A - The assay method of squalene content in a kind of sorghum - Google Patents
The assay method of squalene content in a kind of sorghum Download PDFInfo
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Abstract
The invention discloses a kind of methods of squalene content in measurement sorghum to establish the method that liquid-liquid micro-extraction combination gas chromatography combined with mass spectrometry technology (GC MS) measures squalene content in sorghum by crushing, extraction, extraction.This method is extracted using NaOH ethanol waters, can be reduced the interference of grease in sorghum, be greatly improved the response of object, have the advantages that it is sensitive, fast and accurately, to realize the Accurate Determining of squalene content in sorghum.
Description
Technical field
The invention belongs to analytical chemistry research fields, and in particular to the assay method of squalene content in a kind of sorghum.
Background technology
Squalene is a kind of biologically active terpene substances, has the internal superoxide dismutase (SOD) of raising living
Property, enhancing body immunity, anti-aging, the different physiological roles such as antifatigue, antitumor.Research in recent years is pointed out to pass through meals
The squalene that tonic is filled equally has health efficacy, while also gradually being confirmed by the skin care effect of meal supplement squalene.
Squalene except in shark liver content it is higher in addition to, content is also relatively in the vegetable oil such as olive oil, rice bran oil
Height, although studies have reported that being measured to the squalene content in rice using soxhlet extraction, the spiny dogfish in sorghum
Alkene content is low, and fat content height (3~5%), there has been no the researchs of squalene in sorghum at present.Squalene content in solid sample
Measurement generally use soxhlet extraction enrichment liposoluble constituent after analyzed, for the high sample of fat content such as vegetable oil
Deng the interference for further using saponification to eliminate aliphatic acid etc. then being needed, to improve the instrument response of squalene.These methods
Consumption of organic solvent it is big, the reaction time is long, cumbersome, testing cost is high.
Invention content
In order to solve the above problem of the existing technology, the present invention proposes a kind of measurement side of squalene content in sorghum
Method.
The purpose of the present invention is to provide squalene contents in a kind of sorghum easy, consumption of organic solvent is small, at low cost
Assay method.
The present invention extracts using ethanol water and is added appropriate NaOH, utilizes squalene alcohol-soluble and non-saponified spy
Property, so that squalene is enriched in organic phase by liquid-liquid micro-extraction, reduce the interference of grease in sorghum, improves sorghum
The detection limit and accuracy of middle squalene.Meanwhile operating procedure is easy, greatly reduces the usage amount of organic solvent, saves
The pre-treatment time.
It is realized particular by following technical scheme：
The present invention provides it is a kind of measurement sorghum in squalene content method, be by sorghum seed through crushing, immersion, from
The heart, extraction processing obtain organic phase, are analyzed using gas chromatograph-mass spectrometer (GC-MS) (GC-MS), specifically include following steps：
(1) it crushes：
It is ground using whirlwind pulverizer after sorghum seed removal of impurities as preferred embodiment.
(2) it extracts：
Sample after being crushed as leaching liquor extraction using NaOH ethanol waters, it is spare that centrifugation obtains supernatant.
In step (2), the leaching liquor is NaOH ethanol waters.In the NaOH ethanol waters,
A concentration of 0~0.3mol/L of NaOH；The volume fraction of ethyl alcohol is 0~40%.As preferred embodiment, the concentration of NaOH
For 0.1~0.25mol/L；The volume fraction of ethyl alcohol is 10~20%；Embodiment more preferably, NaOH's is a concentration of
0.15mol/L；The volume fraction of ethyl alcohol is 10%.Crush the quality of sample and leaching liquor (NaOH ethanol waters)：Volume ratio
It is 1:(5~30)；Preferably 1：20.As preferred embodiment, extracted by the way of oscillation.Further, it soaks
It is 5~30min to carry the time, and the revolution of oscillation is 150~450r/min；It is preferred that extraction time is 15min；The revolution of oscillation is
450r/min.Wherein, centrifuge RPMs are 3000~6000r/min, and the time is 5~30min；As preferred embodiment, from
Heart revolution is 4500r/min, time 5min.
(3) it extracts：
It is to be analyzed that upper organic phase is obtained by NaCl, supernatant and extractant mixing, after stratification.
In step (3), the extractant is at least one of pentane, n-hexane or ether.Implement as preferred
Mode, the extractant are the mixture of ether and pentane；It is highly preferred that the volume ratio of ether and pentane is 1:1.As excellent
The embodiment of choosing, in step (3), the mixing by the way of vortex oscillation；Further, duration of oscillation is 1~5min；It is more excellent
It is selected as 1min.The volume ratio of extractant and supernatant is 1:(1~10)；Preferably 1:5.The mass volume ratio of NaCl and supernatant
For (1~5):1；Preferably 3.6：1.
(4) foundation of standard curve：
Squalene is diluted with NaOH- ethanol waters, is configured to a series of squalene standard solution of concentration gradients, successively
The standard solution of each concentration gradient is extracted according to step (3) and obtains organic phase, and is detected using GC-MS.With angle after detection
Squalene concentration is abscissa, and corresponding peak area is that ordinate establishes standard curve.
(5) the GC-MS detections of sample to be tested：
The standard curve comparative analysis that Instrumental results and step (4) are obtained, you can obtain the squalene in sorghum
Content.
Wherein, in step (3) and step (4), the chromatographic condition of GC-MS detections is：1 μ L of sample size, injector temperature 230
DEG C, Splitless injecting samples, capillary chromatographic column is J＆WDB-FFAP quartz capillary column (30m × 0.25mm × 0.25 μm), carrier gas
(He) flow velocity 1mL/min；The Elevated Temperature Conditions of gas-chromatography are：140 DEG C of initial temperature is not kept, according to the rate liter of 15 DEG C/min
Temperature keeps 20min to 230 DEG C；
GC-MS detection Mass Spectrometry Conditions be：Electron ionization sources (Electron ionization, EI)；Electron bombardment energy
Amount：70eV；Ion source temperature：230℃；Quantitative, the quota ion 69 using SIM patterns.
Compared with prior art, technique effect of the invention is embodied in：
1. the present invention has carried out quantitative analysis to the squalene in sorghum for the first time, enriches sorghum micro constitutent and constitute and seek
Support the research of value.The present invention is carried out at the same time by the side saponification and object extraction of establishing squalene in sorghum for the first time, is extracted
The processing method of concentration, and the conditions such as extracting solution type, extractant type are optimized, form NaOH- alcohol-waters
Solution extraction, organic solvent extraction combine the side of squalene content in Gas chromatographyMass spectrometry (GC-MS) measurement sorghum
Method.
2. compared with prior art, the NaOH- ethanol-water solutions of squalene extract, have in sorghum according to the present invention
Solvent extraction processing method, have the advantages that it is easy to operate, quick, economical, accurate, environmentally friendly, specifically compare such as 1 institute of table
Show.
Table 1
| Method | Whether need to heat | Used time (h) | Ethanol consumption (mL) | Extraction efficiency |
| The present invention | It is no | 0.4 | 2 | 91.4% |
| Soxhlet extraction | It is | 5 | 50 | 96.0% |
| Ultrasonic extraction | It is no | 0.4 | 5 | 57.9% |
| Mechanical shaking extraction method | It is no | 2 | 5 | 58.6% |
3, in the present invention detection of squalene be limited to 0.25 μ g/g, be quantitatively limited to 0.82 μ g/g, relative standard deviation is less than
5%, it disclosure satisfy that the quantitative analysis of micro squalene in sorghum.
This method has good linear relationship (R within the measurement range2>0.99) concentration when, with 3 times of signal-to-noise ratio calculates
The detection limit (LOD) of squalene, concentration when with 10 times of signal-to-noise ratio calculate quantitative limit (LOQ), the mark of squalene are listed in table 2
Directrix curve equation, the range of linearity, detection limit, quantitative limit etc..
Table 2
Description of the drawings
Fig. 1 is the influence of squalene content assaying method extracting solution different ethanol concentration in sorghum in the present invention.
Fig. 2 is the mass spectrum of squalene content assaying method extracting solution concentration of alcohol 10% and 20% in sorghum in the present invention
Figure overlapping comparison.
Fig. 3 is the influence of squalene content assaying method extracting solution difference NaOH concentration in sorghum in the present invention.
Fig. 4 is the influence of squalene content assaying method extractant type in sorghum in the present invention.
Fig. 5 is that squalene content assaying method adds the weight of the mass spectrogram before and after squalene standard items in sorghum in the present invention
It is folded to compare.
Specific implementation mode
It is limited with reference to specific embodiment technical scheme of the present invention is further, but claimed model
It encloses and is described made by being not only limited to.
Embodiment 1
A method of squalene content in measurement sorghum is by sorghum seed through crushing, immersion, centrifugation, extraction processing
Organic phase is obtained, is analyzed using gas chromatograph-mass spectrometer (GC-MS) (GC-MS), specifically includes following steps：
(1) it crushes：It is ground with whirlwind pulverizer after sorghum is cleaned spare.
(2) it extracts：Powder sample 1g is weighed in centrifuge tube, is added the NaOH aqueous solutions of 20mL, under the conditions of 450r/min
20min is vibrated, 10min is centrifuged with 5500r/min, supernatant is spare.
(3) it extracts：4g NaCl are weighed in sample bottle, addition 12mL supernatants and 2mL extractants, vortex oscillation 1min,
It is to be analyzed that upper organic phase is obtained after stratification.
(4) it tests and analyzes：By 1 μ L of organic phase to be measured into gas chromatographic analysis, 230 DEG C of injector temperature, Splitless injecting samples,
Capillary chromatographic column is J＆W DB-FFAP quartz capillary columns (30m × 0.25mm × 0.25 μm), carrier gas (He) flow velocity 1mL/min；
The Elevated Temperature Conditions of gas-chromatography are：140 DEG C of initial temperature is not kept, and is warming up to 230 DEG C according to the rate of 15 DEG C/min, is kept
20min；Mass Spectrometry Conditions are：Electron ionization sources (Electron ionization, EI)；Electron bombardment energy：70eV；Ion source
Temperature：230℃；Quantitative, the quota ion 69 using SIM patterns.
A concentration of 0.10mol/L of the NaOH solution.
The extractant is volume ratio 1:1 ether, pentane admixture.
Embodiment 2
A method of squalene content in measurement sorghum is by sorghum seed through crushing, immersion, centrifugation, extraction processing
Organic phase is obtained, is analyzed using gas chromatograph-mass spectrometer (GC-MS) (GC-MS), specifically includes following steps：
(1) it crushes：It is ground with whirlwind pulverizer after sorghum is cleaned spare.
(2) it extracts：Powder sample 1g is weighed in centrifuge tube, the NaOH ethanol waters of 20mL, 450r/min items is added
15min is vibrated under part, 5min is centrifuged with 4500r/min, supernatant is spare.
(3) it extracts：3.6g NaCl are weighed in sample bottle, 10mL supernatants and 2mL extractants, vortex oscillation is added
It is to be analyzed to obtain upper organic phase after stratification by 1min.
(4) it tests and analyzes：By 1 μ L of organic phase to be measured into gas chromatographic analysis, 230 DEG C of injector temperature, Splitless injecting samples,
Capillary chromatographic column is J＆W DB-FFAP quartz capillary columns (30m × 0.25mm × 0.25 μm), carrier gas (He) flow velocity 1mL/min；
The Elevated Temperature Conditions of gas-chromatography are：140 DEG C of initial temperature is not kept, and is warming up to 230 DEG C according to the rate of 15 DEG C/min, is kept
20min；Mass Spectrometry Conditions are：Electron ionization sources (Electron ionization, EI)；Electron bombardment energy：70eV；Ion source
Temperature：230℃；Quantitative, the quota ion 69 using SIM patterns.
A concentration of 0.15mol/L of the NaOH solution.
Volume fraction of ethanol is 10% in the NaOH ethanol solutions.
The extractant is volume ratio 1:1 ether, pentane admixture.
Embodiment 3
A method of squalene content in measurement sorghum is by sorghum seed through crushing, immersion, centrifugation, extraction processing
Organic phase is obtained, is analyzed using gas chromatograph-mass spectrometer (GC-MS) (GC-MS), specifically includes following steps：
(1) it crushes：It is ground with whirlwind pulverizer after sorghum is cleaned spare.
(2) it extracts：Powder sample 1g is weighed in centrifuge tube, the NaOH ethanol solutions of 30mL, 1000r/min conditions is added
Lower oscillation 15min centrifuges 20min with 3500r/min, and supernatant is spare.
(3) it extracts：2g NaCl are weighed in sample bottle, addition 10mL supernatants and 2mL extractants, vortex oscillation 1min,
It is to be analyzed that upper organic phase is obtained after stratification.
(4) it tests and analyzes：By 1 μ L of organic phase to be measured into gas chromatographic analysis, 230 DEG C of injector temperature, Splitless injecting samples,
Capillary chromatographic column is J＆W DB-FFAP quartz capillary columns (30m × 0.25mm × 0.25 μm), carrier gas (He) flow velocity 1mL/min；
The Elevated Temperature Conditions of gas-chromatography are：140 DEG C of initial temperature is not kept, and is warming up to 230 DEG C according to the rate of 15 DEG C/min, is kept
20min；Mass Spectrometry Conditions are：Electron ionization sources (Electron ionization, EI)；Electron bombardment energy：70eV；Ion source
Temperature：230℃；Quantitative, the quota ion 69 using SIM patterns.
A concentration of 0.10mol/L of the NaOH solution.
Volume fraction of ethanol is 20% in the NaOH ethanol solutions.
The extractant is volume ratio 1:1 ether, pentane admixture.
Embodiment 4
A method of squalene content in measurement sorghum is by sorghum seed through crushing, immersion, centrifugation, extraction processing
Organic phase is obtained, is analyzed using gas chromatograph-mass spectrometer (GC-MS) (GC-MS), specifically includes following steps：
(1) it crushes：It is ground with the pulverizer of whirlwind after sorghum is cleaned spare.
(2) it extracts：Powder sample 1g is weighed in centrifuge tube, the NaOH ethanol solutions of 20mL, 450r/min conditions is added
Lower oscillation 15min centrifuges 5min with 4500r/min, and supernatant is spare.
(3) it extracts：3.6g NaCl are weighed in sample bottle, 10mL supernatants and 2mL extractants, vortex oscillation is added
It is to be analyzed to obtain upper organic phase after stratification by 1min.
(4) it tests and analyzes：By 1 μ L of organic phase to be measured into gas chromatographic analysis, 230 DEG C of injector temperature, Splitless injecting samples,
Capillary chromatographic column is J＆W DB-FFAP quartz capillary columns (30m × 0.25mm × 0.25 μm), carrier gas (He) flow velocity 1mL/min；
The Elevated Temperature Conditions of gas-chromatography are：140 DEG C of initial temperature is not kept, and is warming up to 230 DEG C according to the rate of 15 DEG C/min, is kept
20min；Mass Spectrometry Conditions are：Electron ionization sources (Electron ionization, EI)；Electron bombardment energy：70eV；Ion source
Temperature：230℃；Quantitative, the quota ion 69 using SIM patterns.
A concentration of 0.10mol/L of the NaOH solution.
Volume fraction of ethanol is 40% in the NaOH ethanol solutions.
The extractant is volume ratio 1:1 ether, pentane admixture.
Influence of the different volume fraction of ethanol of embodiment 5 to analytical effect
Experiment is used containing NaOH (0.1mol/L) ethyl alcohol (volume fraction 0~40%) aqueous solution in sorghum powder sample respectively
Squalene carry out leaching step.As shown in Figure 1, the concentration of alcohol in extracting solution can impact analysis effect.Squalene is fat
The extracting effect of soluble substance, aqueous solution is poor, therefore tests and carry out extraction behaviour using the ethanol solution of volume fraction 0~40%
Make.Remaining parameter：(1) sorghum sample comminution；(2) sorghum powder sample 1g is weighed in centrifuge tube, and the NaOH second of 20mL is added
Alcoholic solution vibrates 15min under the conditions of 450r/min, centrifuges 5min with 4500r/min, supernatant is spare；(3) it extracts：It weighs
Ether pentane admixture (the volume ratio 1 of 10mL supernatants and 2mL is added in sample bottle in 3.6g NaCl:1) make extractant, whirlpool
1min is vibrated in rotation, and it is to be analyzed to obtain upper organic phase after stratification；(4) it tests and analyzes：By 1 μ L of organic phase to be measured into gas phase color
Spectrum analysis, 230 DEG C of injector temperature, Splitless injecting samples, capillary chromatographic column be J＆W DB-FFAP quartz capillary column (30m ×
0.25mm × 0.25 μm), carrier gas (He) flow velocity 1mL/min；The Elevated Temperature Conditions of gas-chromatography are：140 DEG C of initial temperature is not kept,
230 DEG C are warming up to according to the rate of 15 DEG C/min, keeps 20min；Mass Spectrometry Conditions are：Electron ionization sources (Electron
ionization,EI)；Electron bombardment energy：70eV；Ion source temperature：230℃；Quantitative using SIM patterns, quota ion is
69。
As shown in Figure 1, when volume fraction of ethanol is 10%, the squalene content in leaching liquor is carried than aqueous solution
It is high；When volume fraction of ethanol is increased to 40%, the ethanol content extracted in organic phase increases, and organic phase volume increases, and causes
The concentration of squalene reduces instead；When volume fraction of ethanol is 20%, extraction and extraction results are best.Volume fraction of ethanol 20%
Under the conditions of object squalene 10% ethyl alcohol of response ratio under the conditions of slightly increase, but the aliphatic acid based on oleic acid and linoleic acid
Response is 10% 2 times or more, sees Fig. 2.Consider object response and aliphatic acid disturbed condition, the present invention finally selectes leaching
It is 10% to carry volume fraction of ethanol in solution.
Influence of the concentration of the different NaOH of embodiment 6 to analytical effect
Experiment is respectively with ethyl alcohol (volume fraction 10%) aqueous solution containing NaOH (0~0.3mol/L) to sorghum powder sample
In squalene carry out leaching step.Remaining parameter：(1) sorghum sample comminution；(2) sorghum powder sample 1g is weighed in centrifuge tube
In, the NaOH ethanol solutions of 20mL are added, vibrate 15min under the conditions of 450r/min, 5min, supernatant are centrifuged with 4500r/min
It is spare；(3) it extracts：3.6g NaCl are weighed in sample bottle, the ether pentane admixture (volume of 10mL supernatants and 2mL is added
Than 1:1) make extractant, it is to be analyzed to obtain upper organic phase after stratification by vortex oscillation 1min；(4) it tests and analyzes：It will be to be measured
1 μ L of organic phase are into gas chromatographic analysis, 230 DEG C of injector temperature, Splitless injecting samples, and capillary chromatographic column is J＆W DB-FFAP
Quartzy capillary column (30m × 0.25mm × 0.25 μm), carrier gas (He) flow velocity 1mL/min；The Elevated Temperature Conditions of gas-chromatography are：Starting
140 DEG C of temperature is not kept, and is warming up to 230 DEG C according to the rate of 15 DEG C/min, is kept 20min；Mass Spectrometry Conditions are：Electron ionization sources
(Electronionization,EI)；Electron bombardment energy：70eV；Ion source temperature：230℃；It is quantitative using SIM patterns, it is fixed
It is 69 to measure ion.
As shown in figure 3, the concentration of NaOH can impact analysis effect in extracting solution.When not adding NaOH, fat content mistake
Height can generate pollution to chromatographic apparatus, and object squalene chromatographic peak profile is poor, and response is low.After addition NaOH makes fat saponification, angle
The instrument response of squalene greatly improves, when NaOH concentration is increased to 0.3mol/L, the starch under strong alkali environment in sorghum sample
Gelatinization, system viscosity increased dramatically, and extracting efficiency is reduced and is layered after being unfavorable for extraction.Test determining highly preferred NaOH
A concentration of 0.15mol/L, is as a result shown in Fig. 3.
Influence of the different extractant of embodiment 7 to analytical effect
The type of extractant can also influence effect of extracting.Experiment compares pentane, n-hexane and ether pentane admixture
The effect of extracting of squalene, remaining parameter (1) sorghum sample comminution；(2) sorghum powder sample 1g is weighed in centrifuge tube, is added
Enter the ethanol solution (volume fraction of ethanol 10%) of 20mL 0.15mol/L containing NaOH, vibrates 15min under the conditions of 450r/min, with
4500r/min centrifuges 5min, and supernatant is spare；(3) it extracts：3.6g NaCl are weighed in sample bottle, be added 10mL supernatants and
It is to be analyzed to obtain upper organic phase after stratification by the extractant of 2mL, vortex oscillation 1min；(4) it tests and analyzes：Have to be measured
1 μ L of machine phase are into gas chromatographic analysis, 230 DEG C of injector temperature, Splitless injecting samples, and capillary chromatographic column is J＆W DB-FFAP stones
English capillary column (30m × 0.25mm × 0.25 μm), carrier gas (He) flow velocity 1mL/min；The Elevated Temperature Conditions of gas-chromatography are：Starting temperature
140 DEG C of degree is not kept, and is warming up to 230 DEG C according to the rate of 15 DEG C/min, is kept 20min；Mass Spectrometry Conditions are：Electron ionization sources
(Electronionization,EI)；Electron bombardment energy：70eV；Ion source temperature：230℃；It is quantitative using SIM patterns, it is fixed
It is 69 to measure ion.
The result shows that using ether pentane admixture (volume ratio 1:1) effect of extracting is best, as shown in Figure 4.
Embodiment 8
Further to confirm that the applicability of the present invention, inventor have chosen the sorghum of 6 kinds, be respectively from different
Sorghum plantation area, their squalene content is analyzed using the method for above-mentioned determining condition, specific experiment condition is the same as implementation
Example 2.Relative standard deviation RSD is calculated, as a result such as table 3.
Table 3
From table 3 it can be seen that this method is reproducible, squalene content in different cultivars sorghum can be widely used in
Detection.
Claims (9)
1. a kind of method measuring squalene content in sorghum, it is characterised in that include the following steps：
(1) it crushes；
(2) it extracts：Sample after being crushed as leaching liquor extraction using NaOH ethanol waters, it is spare that centrifugation obtains supernatant；
(3) it extracts：It is to be analyzed that upper organic phase is obtained by NaCl, supernatant and extractant mixing, after stratification；
(4) foundation of standard curve：Squalene is diluted with NaOH- ethanol waters, is configured to a series of spiny dogfish of concentration gradients
Alkene standard solution extracts the standard solution of each concentration gradient according to step (3) obtain organic phase successively, and uses GC-MS
Detection；
(5) the GC-MS detections of sample to be tested.
2. according to the method described in claim 1, it is characterized in that, in step (2), in the NaOH ethanol waters,
A concentration of 0~0.3mol/L of NaOH；The volume fraction of ethyl alcohol is 0~40%；Preferably, a concentration of the 0.1 of NaOH~
0.25mol/L；The volume fraction of ethyl alcohol is 10~20%；It is highly preferred that a concentration of 0.15mol/L of NaOH；The volume of ethyl alcohol
Score is 10%.
3. according to the method described in claim 1, it is characterized in that, in step (2)；The crushing sample and NaOH ethanol waters
The quality of solution：Volume ratio is 1:5~30；Preferably 1：20.
4. according to the method described in claim 1, it is characterized in that, in step (2), extracted by the way of oscillation；Preferably,
The revolution of oscillation is 150~450r/min, and duration of oscillation is 5~30min；It is highly preferred that the revolution of oscillation is 450r/min, shake
It is 15min to swing the time.
5. according to the method described in claim 1, it is characterized in that, in step (2), centrifuge RPMs are 3000~6000r/min,
Time is 5~30min；It is highly preferred that centrifuge RPMs are 4500r/min, time 5min.
6. according to the method described in claim 1, it is characterized in that, in step (3), the extractant is pentane, n-hexane
Or at least one of ether；
Preferably, the extractant is the mixture of ether and pentane；It is highly preferred that the volume ratio of ether and pentane is 1:1.
7. according to the method described in claim 1, it is characterized in that, in step (3), the mixing by the way of vortex oscillation；
Preferably, duration of oscillation is 1~5min；More preferably 1min；
Preferably, in step (3), the volume ratio of extractant and supernatant is 1:1~10；Preferably 1:5.
8. according to the method described in claim 1, it is characterized in that, in step (3), the quality of NaCl and supernatant：Volume ratio
It is 1~5:1；Preferably 3.6：1.
9. according to the method described in claim 1, it is characterized in that, in step (3) and step (4), the chromatostrip of GC-MS detections
Part is：1 μ L of sample size, 230 DEG C of injector temperature, Splitless injecting samples, capillary chromatographic column are J＆W DB-FFAP quartz capillary columns
30m × 0.25mm × 0.25 μm, carrier gas He flow velocitys 1mL/min；The Elevated Temperature Conditions of gas-chromatography are：140 DEG C of initial temperature is not protected
It holds, is warming up to 230 DEG C according to the rate of 15 DEG C/min, keeps 20min；
GC-MS detection Mass Spectrometry Conditions be：Electron ionization sources；Electron bombardment energy：70eV；Ion source temperature：230℃；Using
SIM patterns are quantitative, quota ion 69.
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