CN109613161A - A kind of β based on headspace gas chromatography-carypohyllene content assaying method - Google Patents
A kind of β based on headspace gas chromatography-carypohyllene content assaying method Download PDFInfo
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- CN109613161A CN109613161A CN201811607065.7A CN201811607065A CN109613161A CN 109613161 A CN109613161 A CN 109613161A CN 201811607065 A CN201811607065 A CN 201811607065A CN 109613161 A CN109613161 A CN 109613161A
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Abstract
The invention discloses a kind of β based on headspace gas chromatography-carypohyllene content assaying method, its principle is to extract β-carypohyllene first from sample, then β-carypohyllene is injected in head-space sampler, top tank air is extracted when reaching vapor liquid equilibrium, it is separated using gas-chromatography, hydrogen flame ionization detector detects β-carypohyllene concentration, finally calculates β-carypohyllene content with this.Headspace gas chromatography precision of the present invention is high, can detecte the content of ppm rank, and detection sensitivity is high.Detection method Sample pretreatment process of the invention is simple to operation.The determination step of detection method of the invention is all automation equipment operation, can continuously detect samples up to a hundred, reads accurate data, each sample test time is short.For detection method of the invention relative to existing gas chromatography-mass spectrometry and capillary gas chromatography, price advantage is obvious, and required reagent is nontoxic, environmentally protective.
Description
Technical field
The invention belongs to life sciences, and in particular to a kind of β-carypohyllene based on headspace gas chromatography is containing measuring
Determine method.
Background technique
Fragrance is the important component of flavor quality, and many plants and fruit all have fragrance.As market is to fruit
The increase that the requirement of quality is higher and higher and grocery trade, cosmetic industry are to natural flavor material requisite, research plant (including water
Fruit) aroma substance has become an emerging subject.Volatile materials is the main source of fragrance, along with plant (including water
Fruit) maturation, the type and content of volatile materials can all generate certain variation.
β-carypohyllene is mainly used for preparing smart Counterfeit Item and fixastive, belongs to natural equivalent fragrance and artificial perfume, can be by
It is isolated in clove leaf oil, clove-stems oil, cinnamon leaves oil etc., meanwhile, β-carypohyllene has anticancer, anti-hypertension and anti-inflammatory
The function such as analgesia.Existing detection β-carypohyllene content method mainly has capillary gas chromatography and gas phase color in document at present
Spectrum-Mass Spectrometry.
Capillary gas chromatography one carrys out extraction process complexity, and extracting solution is mostly organic solvent；Two need internal standard is added
Object；Three carry out sensitivity and detect limit all to need to be continued to improve.Gas chromatography-mass spectrometry is expensive, and to environment, examination
The requirements such as agent, instrument and equipment are very high.With the rapid development of chromatographic technique in recent years, applied in the detection of fragrance substance
It is more and more extensive.It is generally cumbersome for the pre-treatment of sample but now on the market based on the detection technique of gas chromatography.
Headspace gas chromatography (HS-GC) is also known as saturated slarry analysis, is a kind of joint operation technology, generallys use
Sample introduction needle carries out extraction absorption to solid, liquid, gas etc. under certain temperature under certain condition, then in gas chromatographic analysis
Desorption injection is carried out on instrument.Standard gas technology is with its simple and practical advantage in environment measuring (volatility halogen in such as drinking water
For volatile organic matter in hydrocarbon and trade effluent), organic residual solvent detection in drug, food, forensic science, petrochemical industry,
The fields such as packaging material, coating and wine-making industry analysis are widely used.
Summary of the invention
In order to meet the demand, the present invention is intended to provide a kind of β-carypohyllene based on headspace gas chromatography is containing measurement
Determine method, has detection accurate high, high sensitivity, Sample pretreatment is convenient, and continuity is good, and detection speed is fast, and cost is relatively low, ring
Protect the advantages that nontoxic.
To realize above-mentioned technical purpose and the technique effect, the invention is realized by the following technical scheme:
A kind of β based on headspace gas chromatography-carypohyllene assay kit, including following reagent:
Reagent one, ultrapure water 10mL × 1 bottle, 4 DEG C of preservations；
Reagent two, quasi- product 0.1mL × 1 of β-carypohyllene, -20 DEG C of preservations.
A kind of β based on headspace gas chromatography-carypohyllene content assaying method, principle are to infuse in head-space sampler
Enter β-carypohyllene, top tank air is extracted when reaching vapor liquid equilibrium, is separated using gas-chromatography, hydrogen flame ionization detector detection
β-carypohyllene concentration calculates β-carypohyllene content with this.
The present invention detects β-carypohyllene content using headspace gas chromatography for the first time, the specific steps of which are as follows:
The preparation of step 1 instrument and articles；
Prepare gas chromatograph, 30m KB-624 pillar, hydrogen flame ionization detector, head-space sampler, 20mL band gasket glass
Container (50), ultrapure water, the quasi- product of β-carypohyllene；
The extraction of step 2 β-carypohyllene；
Tissue samples: weighing 0.5g tissue samples, and record quality, grind, and is put into 20mL band gasket glass container, is added
2mL ultrapure water, is stoppered rubber stopper, and vapor liquid equilibrium is reached on head-space sampler, is balancing after a certain period of time, is taking top tank air,
Inject gas chromatographic detection；
Liquid sample: take 2mL liquid sample to be put into 20mL band gasket glass container (can dilute) according to the actual situation, in head space
Reach vapor liquid equilibrium on sample injector, is balancing after a certain period of time, taking top tank air, injecting gas chromatographic detection；
The preparation of step 3 β-caryophyllene standard product:
Using β-caryophyllene standard product and ultrapure water, being configured to concentration respectively is 50ppm, 10ppm, 5ppm, 1ppm, 0.5ppm,
The β of 0.1ppm-caryophyllene standard product solution；
The operation of step 4 β-carypohyllene assay；
1) computer, carrier gas, air source generator, hydrogen generator, gas phase host are opened, software is opened, is arranged in method group
Injector temperature, detector temperature, column oven temperature, and temperature program is set, store method group that setting completed, and delivery method
To instrument；
2) the ml headspace bottle temperature of head-space sampler, quantitative loop temperature, transmission line temperature and equilibration time are set；
3) when three temperature of instrument all reach setting value, instrument igniting is carried out；After lighting a fire successfully, start to monitor baseline, to base
After the steady certain time of line, start sample introduction；
4) β-caryophyllene standard product gas 1mL is injected, β-carypohyllene is separated in 20min, and in β-carypohyllene retention time
Place's (pillar is different, and retention time is variant), calculates β-caryophyllene standard product peak area of various concentration；
5) β-carypohyllene sample gas 1mL is injected, β-carypohyllene sample peak area is detected at corresponding retention time；
The calculating of step 5 β-carypohyllene content；
With β-caryophyllene standard product concentration (ppm) for abscissa, β-caryophyllene standard product peak area is that ordinate calculates β-China pink
Alkene standard curve；
β-carypohyllene sample peak area is substituted into standard curve, calculates β-carypohyllene concentration in β-carypohyllene sample；
Calculated β-carypohyllene concentration obtains β-carypohyllene and contains divided by the quality of tissue samples or the volume of liquid sample
Amount.
Further, in step 1, the storage temperature of ultrapure water is 4 DEG C, and β-caryophyllene standard product storage temperature is -20
℃。
Further, in step 2, when extracting β-carypohyllene, the glass container reaches gas on the head-space sampler
The time of liquid balance is 20min.
Further, in step 3, the β-caryophyllene standard product concrete operations for preparing various concentration are as follows:
10 μ L β-carypohyllene is taken, is added in 10mL ultrapure water, it is molten to be configured to β-caryophyllene standard product that concentration is 1000ppm
Liquid, then diluted with ultrapure water, being configured to concentration respectively is 50ppm, 10ppm, 5ppm, 1ppm, 0.5ppm, β-stone of 0.1ppm
Bamboo alkene standard solution.
Further, the 1 of step 4) in, the injector temperature being arranged in method group is 250 DEG C, and detector temperature is
250 DEG C, column oven temperature is 60 DEG C, and temperature program is arranged according to the form below:
。
Further, the 2 of step 4) in, the ml headspace bottle temperature that head-space sampler is arranged is 85 DEG C, and quantitative loop temperature is 95
DEG C, transmission line temperature is 105 DEG C, equilibration time 20min.
Further, the 3 of step 4) in, the baseline stable time is 30min.
Further, the 4 of step 4) in, β-carypohyllene retention time is in 18.346min or so.
Compared with prior art, the beneficial effects of the present invention are:
1, headspace gas chromatography precision used by detection method of the invention is high, can detecte the content of ppm rank,
Detection sensitivity is high.
2, detection method Sample pretreatment process of the invention is simple to operation.
3, the determination step of detection method of the invention is all automation equipment operation, can continuously detect samples up to a hundred,
Accurate data is read, each sample test time is short.
4, detection method of the invention is relative to existing gas chromatography-mass spectrometry and capillary gas chromatography,
Price advantage is obvious, and required reagent is nontoxic, environmentally protective.
The above description is only an overview of the technical scheme of the present invention, in order to better understand the technical means of the present invention,
And can be implemented in accordance with the contents of the specification, it is described in detail below with presently preferred embodiments of the present invention.Specific reality of the invention
Mode is applied to be shown in detail by following embodiment.
Specific embodiment
Below in conjunction with embodiment, next the present invention will be described in detail.
A kind of β based on headspace gas chromatography-carypohyllene assay kit, including following reagent:
Reagent one, ultrapure water 10mL × 1 bottle, 4 DEG C of preservations；
Reagent two, quasi- product 0.1mL × 1 of β-carypohyllene, -20 DEG C of preservations.
A kind of β based on headspace gas chromatography-carypohyllene content assaying method, principle are to infuse in head-space sampler
Enter β-carypohyllene, top tank air is extracted when reaching vapor liquid equilibrium, is separated using gas-chromatography, hydrogen flame ionization detector detection
β-carypohyllene concentration calculates β-carypohyllene content with this.
The present invention detects β-carypohyllene content using headspace gas chromatography for the first time, the specific steps of which are as follows:
The preparation of step 1 instrument and articles；
Prepare gas chromatograph, 30m KB-624 pillar, hydrogen flame ionization detector, head-space sampler, 20mL band gasket glass
Container (50), the ultrapure water of 4 DEG C of preservations, the quasi- product of β-carypohyllene of -20 DEG C of preservations；
The extraction of step 2 β-carypohyllene；
Tissue samples: weighing 0.5g tissue samples, and record quality, grind, and is put into 20mL band gasket glass container, is added
2mL ultrapure water, is stoppered rubber stopper, and vapor liquid equilibrium is reached on head-space sampler, and equilibration time 20min takes top tank air, injection
Gas chromatographic detection；
Liquid sample: take 2mL liquid sample to be put into 20mL band gasket glass container (can dilute) according to the actual situation, in head space
Reach vapor liquid equilibrium on sample injector, equilibration time 20min takes top tank air, injects gas chromatographic detection；
The preparation of step 3 β-caryophyllene standard product:
10 μ L β-carypohyllene is taken, is added in 10mL ultrapure water, it is molten to be configured to β-caryophyllene standard product that concentration is 1000ppm
Liquid, then diluted with ultrapure water, being configured to concentration respectively is 50ppm, 10ppm, 5ppm, 1ppm, 0.5ppm, β-stone of 0.1ppm
Bamboo alkene standard solution；
The operation of step 4 β-carypohyllene assay；
1) computer, carrier gas, air source generator, hydrogen generator, gas phase host are opened, software is opened, is arranged in method group
250 DEG C of injector temperature, 250 DEG C of detector temperature, column oven temperature 60 C is arranged according to the temperature program in following table, setting
Finish store method group, and delivery method is to instrument；
2) 85 DEG C of ml headspace bottle temperature of setting head-space sampler, 95 DEG C of quantitative loop, 105 DEG C of transmission line temperature, equilibration time
20min；
3) when three temperature of instrument all reach setting value, instrument igniting is carried out；After lighting a fire successfully, start to monitor baseline, to base
After the steady 30min of line, start sample introduction；
4) β-caryophyllene standard product gas 1mL is injected, β-carypohyllene is separated in 20min, β-carypohyllene retention time is
18.346min or so (pillar is different, and retention time is variant), calculates β-caryophyllene standard product peak area of various concentration；
5) β-carypohyllene sample gas 1mL is injected, β-carypohyllene sample peak area is detected at corresponding retention time；
The calculating of step 5 β-carypohyllene content；
With β-caryophyllene standard product concentration (ppm) for abscissa, β-caryophyllene standard product peak area is that ordinate calculates β-China pink
Alkene standard curve；
β-carypohyllene sample peak area is substituted into standard curve, calculates β-carypohyllene concentration in β-carypohyllene sample；
Calculated β-carypohyllene concentration obtains β-carypohyllene and contains divided by the quality of tissue samples or the volume of liquid sample
Amount.
Simply to illustrate that technical concepts and features of the invention, its purpose is allows in the art above-described embodiment
Those of ordinary skill cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all
It is changes or modifications equivalent made by the essence of content according to the present invention, should be covered by the scope of protection of the present invention.
Claims (8)
1. a kind of β based on headspace gas chromatography-carypohyllene content assaying method, which is characterized in that specific step is as follows:
The preparation of step 1 instrument and articles；
Prepare gas chromatograph, 30m KB-624 pillar, hydrogen flame ionization detector, head-space sampler, 20mL band gasket glass
Container, ultrapure water, the quasi- product of β-carypohyllene；
The extraction of step 2 β-carypohyllene；
Tissue samples: weighing 0.5g tissue samples, and record quality, grind, and is put into 20mL band gasket glass container, is added
2mL ultrapure water, is stoppered rubber stopper, and vapor liquid equilibrium is reached on head-space sampler, is balancing after a certain period of time, is taking top tank air,
Inject gas chromatographic detection；
Liquid sample: it takes 2mL liquid sample to be put into 20mL band gasket glass container, it is flat that gas-liquid is reached on head-space sampler
Weighing apparatus is balancing after a certain period of time, is taking top tank air, injecting gas chromatographic detection；
The preparation of step 3 β-caryophyllene standard product:
Using β-caryophyllene standard product and ultrapure water, being configured to concentration respectively is 50ppm, 10ppm, 5ppm, 1ppm, 0.5ppm,
The β of 0.1ppm-caryophyllene standard product solution；
The operation of step 4 β-carypohyllene assay；
1) computer, carrier gas, air source generator, hydrogen generator, gas phase host are opened, software is opened, is arranged in method group
Injector temperature, detector temperature, column oven temperature, and temperature program is set, store method group that setting completed, and delivery method
To instrument；
2) the ml headspace bottle temperature of head-space sampler, quantitative loop temperature, transmission line temperature and equilibration time are set；
3) when three temperature of instrument all reach setting value, instrument igniting is carried out；After lighting a fire successfully, start to monitor baseline, to base
After the steady certain time of line, start sample introduction；
4) β-caryophyllene standard product gas 1mL is injected, β-carypohyllene is separated in 20min, and in β-carypohyllene retention time
Place, calculates β-caryophyllene standard product peak area of various concentration；
5) β-carypohyllene sample gas 1mL is injected, β-carypohyllene sample peak area is detected at corresponding retention time；
The calculating of step 5 β-carypohyllene content；
With β-caryophyllene standard product concentration (ppm) for abscissa, β-caryophyllene standard product peak area is that ordinate calculates β-China pink
Alkene standard curve；
β-carypohyllene sample peak area is substituted into standard curve, calculates β-carypohyllene concentration in β-carypohyllene sample；
Calculated β-carypohyllene concentration obtains β-carypohyllene and contains divided by the quality of tissue samples or the volume of liquid sample
Amount.
2. the β based on headspace gas chromatography-carypohyllene content assaying method according to claim 1, which is characterized in that
In step 1, the storage temperature of ultrapure water is 4 DEG C, and β-caryophyllene standard product storage temperature is -20 DEG C.
3. the β based on headspace gas chromatography-carypohyllene content assaying method according to claim 1, which is characterized in that
In step 2, when extracting β-carypohyllene, the time that the glass container reaches vapor liquid equilibrium on the head-space sampler is
20min。
4. the β based on headspace gas chromatography-carypohyllene content assaying method according to claim 1, which is characterized in that
In step 3, the β-caryophyllene standard product concrete operations for preparing various concentration are as follows:
10 μ L β-carypohyllene is taken, is added in 10mL ultrapure water, it is molten to be configured to β-caryophyllene standard product that concentration is 1000ppm
Liquid, then diluted with ultrapure water, being configured to concentration respectively is 50ppm, 10ppm, 5ppm, 1ppm, 0.5ppm, β-stone of 0.1ppm
Bamboo alkene standard solution.
5. the β based on headspace gas chromatography-carypohyllene content assaying method according to claim 1, which is characterized in that
The 1 of step 4) in, the injector temperature being arranged in method group is 250 DEG C, and detector temperature is 250 DEG C, and column oven temperature is
60 DEG C, and temperature program is arranged according to the form below:
。
6. the β based on headspace gas chromatography-carypohyllene content assaying method according to claim 1, which is characterized in that
The 2 of step 4) in, the ml headspace bottle temperature that head-space sampler is arranged is 85 DEG C, and quantitative loop temperature is 95 DEG C, and transmission line temperature is 105
DEG C, equilibration time 20min.
7. the β based on headspace gas chromatography-carypohyllene content assaying method according to claim 1, which is characterized in that
The 3 of step 4) in, the baseline stable time is 30min.
8. the β based on headspace gas chromatography-carypohyllene content assaying method according to claim 1, which is characterized in that
The 4 of step 4) in, β-carypohyllene retention time is 18.346min.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH1130610A (en) * | 1997-07-09 | 1999-02-02 | Sapporo Breweries Ltd | Hop originated volatile aromatic component measuring method |
| CN104931605A (en) * | 2015-05-20 | 2015-09-23 | 增城华栋调味品有限公司 | Method for detecting quality of pepper powder based on component analysis |
| CN107621496A (en) * | 2016-07-13 | 2018-01-23 | 上海凯宝药业股份有限公司 | A kind of detection method of Chinese medicine composition |
| US20180284145A1 (en) * | 2015-01-26 | 2018-10-04 | Biotech Institute, Llc | Systems, apparatuses, and methods for classification |
-
2018
- 2018-12-27 CN CN201811607065.7A patent/CN109613161A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH1130610A (en) * | 1997-07-09 | 1999-02-02 | Sapporo Breweries Ltd | Hop originated volatile aromatic component measuring method |
| US20180284145A1 (en) * | 2015-01-26 | 2018-10-04 | Biotech Institute, Llc | Systems, apparatuses, and methods for classification |
| CN104931605A (en) * | 2015-05-20 | 2015-09-23 | 增城华栋调味品有限公司 | Method for detecting quality of pepper powder based on component analysis |
| CN107621496A (en) * | 2016-07-13 | 2018-01-23 | 上海凯宝药业股份有限公司 | A kind of detection method of Chinese medicine composition |
Non-Patent Citations (3)
| Title |
|---|
| 李红等: "顶空萃取-气相色谱-质谱法测定一种儿童专用防痱止痒水的挥发性成分", 《现代科学仪器》 * |
| 梁志勤等: "马尾松松脂重质松节油化学组成的研究 ", 《林产化学与工业》 * |
| 耿迪等: "酒花香气物质检测方法的建立及评估 ", 《啤酒科技》 * |
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Application publication date: 20190412 |