CN109212067A - The detection method and test sample processing method of tobacco and tobacco product Fatty Acids - Google Patents
The detection method and test sample processing method of tobacco and tobacco product Fatty Acids Download PDFInfo
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- CN109212067A CN109212067A CN201811107004.4A CN201811107004A CN109212067A CN 109212067 A CN109212067 A CN 109212067A CN 201811107004 A CN201811107004 A CN 201811107004A CN 109212067 A CN109212067 A CN 109212067A
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- tobacco
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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
- G01N30/62—Detectors specially adapted therefor
- G01N30/72—Mass spectrometers
- G01N30/7206—Mass spectrometers interfaced to gas chromatograph
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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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/025—Gas 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
- 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
- G01N2030/067—Preparation by reaction, e.g. derivatising the sample
Abstract
The present invention relates to a kind of tobacco and the detection methods and test sample processing method of tobacco product Fatty Acids, belong to tobacco component detection technique field.The detection method of tobacco and tobacco product Fatty Acids of the invention, includes the following steps: to mix sample with extractant and extracts, and derivatization reagent is then added and performs the derivatization reaction, then progress GC-MS/MS test；The extractant is methylene chloride.The detection method of tobacco and tobacco product Fatty Acids of the invention is extracted using methylene chloride as solvent, and carries out derivative reaction, the 12- methyl tetradecanoic acid being capable of detecting when in tobacco leaf.It moreover, cooperation GC-MS/MS technology, can be realized that sensitivity is higher, detection limit is lower, can be used to analyze the higher fatty acids of the low content in tobacco leaf: undecanoic acid, 12- methyl tetradecanoic acid, heptadecanoic acid, 20 acid.
Description
Technical field
The present invention relates to a kind of tobacco and the detection method and test sample processing method of tobacco product Fatty Acids,
Belong to tobacco component detection technique field.
Background technique
Organic acid is widely present in tobacco, and content is about 10%~16%, and the fragrance and smell to cigarette have important
Effect, the higher fatty acids of C10 or more are wherein very important one kind.Chain saturated fatty acids and unsaturated fatty acid can increase
The fat and wax taste for adding flue gas, make smoke alcohol and adjust the pH value of tobacco, can also increase flue gas concentration, influence flue gas perfume
Gas.Different long chain fatty acids fragrance also have different, for example, undecanoic acid, lauric acid/dodecanoic acid can make smoke alcohol and and suitable；Ten
Tetracid (i.e. myristic acid) has soft flue gas to act on, and with cigarette perfume harmony, improves sweet and mellow flavor；Linolenic acid has ceride gas
Breath, can increase richness, and have certain irritation；Hexadecylic acid is in addition to also having sweet taste, and can increase richness with cured meat and fish；
20 acid have nut taste, can increase flue gas concentration.
Method derived from methanolic is generally used to the detection of tobacco leaf Fatty Acids, but this method is only suitable for content
The measurement of high higher fatty acids, such as hexadecylic acid, stearic acid, stearic acid, linoleic acid.Also there is document report using acceleration solvent
It extracts Silylation and measures higher fatty acids, but since some higher aliphatic acid contents are low, it is not easy to be detected such as 11
Acid, 12- methyl tetradecanoic acid, heptadecanoic acid, 20 acid etc..
Application publication number is that the Chinese invention patent application of CN106353429A discloses a kind of tobacco based on organic acid
The method of discrimination of fresh tobacco leaves sample quality in metabolism group.It uses methylene chloride/acetonitrile to carry out ultrasound as extractant
Extraction, then performs the derivatization reaction with BSTFA, then carry out GC/MS analysis, detects the content of organic acid.This method can be examined
The content ratio of heptadecanoic acid and butyric acid is surveyed, but the lower higher fatty acids of other contents can not be detected.
Summary of the invention
The purpose of the present invention is to provide in a kind of tobacco for being capable of detecting when low content higher fatty acids and tobacco product
The detection method of higher fatty acids.
The object of the invention is also to provide a kind of tobacco and the test sample processing methods of tobacco product.
To achieve the above object, the technical solution of the detection method of tobacco of the invention and tobacco product Fatty Acids
It is:
The detection method of a kind of tobacco and tobacco product Fatty Acids, includes the following steps: sample and extractant
Mixing is extracted, and derivatization reagent is then added and performs the derivatization reaction, then carries out GC-MS/MS test；The extractant
For methylene chloride.
The detection method of tobacco and tobacco product Fatty Acids of the invention is carried out using methylene chloride as solvent
Extraction, and derivative reaction is carried out, it is capable of detecting when the low contents higher fatty acids such as the 12- methyl tetradecanoic acid in tobacco leaf.And
And cooperation GC-MS/MS technology, can be realized sensitivity it is higher, detection limit it is lower, can be used to analyze the low content in tobacco leaf
Higher fatty acids.
The higher fatty acids is undecanoic acid, lauric acid, myristic acid, 12- methyl-tetradecanoic acid, palmitinic acid, pearl
At least one of acid, linoleic acid, oleic acid, linolenic acid, stearic acid, arachidic acid.
Further, method of the invention can measure 11 kinds of higher aliphatics with characteristic perfume and smell in tobacco leaf simultaneously
Acid expands the detection range of higher fatty acids.The described 11 kinds higher fatty acids with characteristic perfume and smell be undecanoic acid,
Lauric acid, myristic acid, 12- methyl-tetradecanoic acid, palmitinic acid, pearl acid, linoleic acid, oleic acid, linolenic acid, stearic acid, peanut
Acid.
The tobacco sample carries out sample treatment before mixing with extractant, and the sample treatment includes: that powder is made in sample
End, sieving.The sieving was 40 meshes.The powder that is made is that sample is pulverized.The dosage of the extractant is every
1g sample corresponds to 6-20mL extractant.
It is the 10-50min that is vortexed after mixing sample with extractant, inner mark solution that the mixing, which carries out extraction,.The vortex
Revolving speed be 2500rpm.
When sample is mixed with extractant, inner mark solution is added.The dosage of the inner mark solution is that every 1g sample corresponds to 50-
100 μ L inner mark solutions.The inner mark solution is nonadecanoic acid solution.The concentration of the inner mark solution is 50-100 μ g/mL.
The tobacco and tobacco product are tobacco leaf or pipe tobacco.
Sample is mixed with extractant extracted after obtain extract liquor.
The derivatization reagent is bis- (trimethylsilyl) trifluoroacetamides (BSTFA).
The dosage of the derivatization reagent is that the extract liquor of every 0.5-1mL corresponds to 50-200 μ L derivatization reagent.
The time of the derivative reaction is 10-40min.
The sample is mixed with extractant extracted after be filtered.Described be filtered into was carried out using organic phase filter membrane
Filter.
The condition of the GC-MS/MS test are as follows: injector temperature is 280 DEG C, and sample volume is 1.0 μ L.Split ratio is 10:
1.Carrier gas is helium.Constant current flow velocity is 1.0mL/min.
Chromatographic column uses DB-5MS chromatographic column.The specification of chromatographic column is 60m × 0.25mmi.d. × 0.25 μm d.f.).It rises
Warm program are as follows: 80 DEG C of initial temperature, rise to 280 DEG C with the heating rate of 10 DEG C/min, keep the temperature 20min.Ionization mode is EI.Transmission line
Temperature is 280 DEG C.Ion source temperature: 300 DEG C.Scanning mode is multiple-reaction monitoring (MRM) mode.CID gas pressure is
2.0mTorr；Monitoring time to each ion is 100ms.
The technical solution of the test sample processing method of tobacco and tobacco product of the invention is:
The test sample processing method of a kind of tobacco and tobacco product, includes the following steps: to mix sample with extractant
Extracted, then be added derivatization reagent perform the derivatization reaction to get；The extractant is methylene chloride.
Detailed description of the invention
Fig. 1 is the chromatogram of undecanoic acid in the standard solution and sample of undecanoic acid in the embodiment of the present invention 1；
Fig. 2 is the chromatography of lauric acid/dodecanoic acid in the standard solution and sample of lauric acid/dodecanoic acid (lauric acid) in the embodiment of the present invention 1
Figure；
Fig. 3 is the chromatography of tetradecylic acid in the standard solution and sample of tetradecylic acid (myristic acid) in the embodiment of the present invention 1
Figure；
Fig. 4 is 12- methyltetradecylphosphine in the standard solution and sample of 12- methyl tetradecanoic acid in the embodiment of the present invention 1
The chromatogram of acid；
Fig. 5 is the chromatogram of hexadecylic acid in the standard solution and sample of hexadecylic acid (palmitinic acid) in the embodiment of the present invention 1；
Fig. 6 is the chromatogram of heptadecanoic acid in the standard solution and sample of heptadecanoic acid in the embodiment of the present invention 1；
Fig. 7 is the standard solution of 1 Linoleic acid of the embodiment of the present invention and the chromatogram of sample Linoleic acid；
Fig. 8 is the chromatogram of oleic acid in the standard solution and sample of oleic acid in the embodiment of the present invention 1；
Fig. 9 is linolenic chromatogram in linolenic standard solution in the embodiment of the present invention 1 and sample；
Figure 10 is stearic chromatogram in stearic standard solution in the embodiment of the present invention 1 and sample；
Figure 11 is the chromatography of 20 acid in the standard solution and sample of 20 sour (arachidic acids) in the embodiment of the present invention 1
Figure；
Figure 12 is the chromatogram of internal standard compound in the standard solution and sample of nonadecanoic acid in the present invention.
Specific embodiment
Technical solution of the present invention is described further combined with specific embodiments below.
The preparation of standard solution:
A certain amount of standard items are weighed respectively in 10mL volumetric flask, prepare single mark mother liquor.By single mark mother liquor dilution, or directly
It takes mother liquor a certain amount of, is added in 10mL volumetric flask, the mixed mark of graticule maximum concentration is prepared, by diluting production standard curve step by step.
Six grades of graticule concentration ranges are as shown in table 1, and unit is μ g/mL.
The titer parameter of each fatty acid of table 1
Embodiment 1
The detection method of the tobacco Fatty Acids of the present embodiment includes the following steps:
1) tobacco leaf C3F sample is clayed into power, crosses 40 meshes and obtains tobacco powder.0.5 gram of tobacco powder is taken to be put into sample bottle, then
5mL methylene chloride is added into sample bottle and makees extractant, ten that 100 μ L concentration are 50 μ g/mL are then added into sample bottle again
Nine sour inner mark solutions.30min is extracted with the velocity vortex of 2500rpm, organic phase filter membrane is crossed, obtains extract liquor.
2) extract liquor 0.5mL obtained in step 1) is taken, bis- (trimethylsilyl) trifluoroacetyls of 150 μ L are added thereto
Amine (BSTFA) derivatization reagent performs the derivatization reaction 10min at room temperature.
3) system after derivative reaction is subjected to GC-MS/MS test.
GC-MS/MS test condition are as follows:
GC-MS/MS instrument injector temperature is 280 DEG C, and sample volume: 1.0 μ L, split ratio 10:1, carrier gas are helium, permanent
Stream flow velocity is 1.0mL/min.
Chromatographic column uses DB-5MS chromatographic column (i.d. × 0.25 μm 60m × 0.25mm d.f.)；Temperature programming: initial temperature 80
DEG C, 280 DEG C are risen to the heating rate of 10 DEG C/min, keeps the temperature 20min.Ionization mode: EI；Transmission line temperature: 280 DEG C；Ion
Source temperature: 300 DEG C.Scanning mode: multiple-reaction monitoring (MRM) mode, CID gas pressure are 2.0mTorr；To each ion
Monitoring time is 100ms.
Impact energy after 11 kinds of higher fatty acids and internal standard the retention time of the compound, parent ion, daughter ion and optimization
The parameters such as amount (CE) value are shown in Table 2.The chromatogram of target compound is as shown in figs. 1-12 in each Fatty acid standards solution and sample.
The mass spectral analysis parameter of each fatty acid of table 2
Embodiment 2
Flue-cured tobacco, burley tobaccos, four kinds of Turkish tobaccos, Dark sun-cured each one of different type tobacco leaf C3F samples are chosen, using embodiment
Method in 1 carries out sample pre-treatments and the analysis of GC-MS/MS instrument.11 kinds of higher fatty acids content results such as 3 institute of table in tobacco leaf
Show.
The content (μ g/g) of 11 kinds of higher fatty acids in 3 different type tobacco leaf of table
Test example
Typical flue-cured tobacco sample is chosen, method is carried out in a few days and day to day precision is tested.In on the same day, to selected sample one
5 parallel laboratory tests are carried out in it to investigate method withinday precision, are tested to investigate method within five days continuous to selected sample
Day to day precision.It is reference with actual sample content, respectively plus 50,100,200 μ L of mixed mark, mixed mark concentration are as shown in table 4.The side of doing
Method accuracy characterization, rate of recovery average result are as shown in table 5.The result shows that: each target acid ingredient average recovery rate exists
85.66%-106.12%, in a few days, day to day precision be respectively less than 10%.
4 mark-on liquid concentration of table
| ug/mL | |
| Undecanoic acid | 2.5 |
| Lauric acid/dodecanoic acid | 2 |
| Tetradecylic acid | 200 |
| 12- methyl tetradecanoic acid | 20 |
| Hexadecylic acid | 5000 |
| Heptadecanoic acid | 200 |
| Linoleic acid | 2000 |
| Oleic acid | 2000 |
| Linolenic acid | 2000 |
| Stearic acid | 2000 |
| 20 acid | 1000 |
Detection limit and quantitative limit are calculated by infinite dilution method, the results showed that, this method sensitivity fully meets test specimens
The needs of product.The results are shown in Table 5.
Precision, the rate of recovery, detection limit and the quantitative limit of the method for the invention of table 5
Claims (10)
1. the detection method of a kind of tobacco and tobacco product Fatty Acids, characterized by the following steps: by sample
It mixes and is extracted with extractant, derivatization reagent is then added and performs the derivatization reaction, then carries out GC-MS/MS test；Institute
Stating extractant is methylene chloride.
2. the detection method of tobacco according to claim 1 and tobacco product Fatty Acids, it is characterised in that: described
The dosage of extractant is that every 1g sample corresponds to 6-20mL extractant.
3. the detection method of tobacco according to claim 1 and tobacco product Fatty Acids, it is characterised in that: by sample
When product are mixed with extractant, it is added inner mark solution, it is molten that the dosage of the inner mark solution is that every 1g sample corresponds to 50-100 μ L internal standard
Liquid.
4. the detection method of tobacco according to claim 3 and tobacco product Fatty Acids, it is characterised in that: described
Inner mark solution is nonadecanoic acid solution.
5. the detection method of tobacco according to claim 3 and tobacco product Fatty Acids, it is characterised in that: described
The concentration of inner mark solution is 50-100 μ g/mL.
6. the detection method of tobacco according to claim 1 and tobacco product Fatty Acids, it is characterised in that: described
Derivatization reagent is bis- (trimethylsilyl) trifluoroacetamides.
7. the detection method of tobacco according to claim 1 and tobacco product Fatty Acids, it is characterised in that: sample
It is mixed with extractant and obtains extract liquor after being extracted；The extract liquor that the dosage of the derivatization reagent is every 0.5-1mL is corresponding
50-200 μ L derivatization reagent.
8. the detection method of tobacco according to claim 1 and tobacco product Fatty Acids, it is characterised in that: described
The time of derivative reaction is 10-40min.
9. the detection method of tobacco according to claim 1 and tobacco product Fatty Acids, it is characterised in that: described
It is the 10-50min that is vortexed after mixing sample with extractant, inner mark solution that mixing, which carries out extraction,.
10. the test sample processing method of a kind of tobacco and tobacco product, characterized by the following steps: by sample with
Extractant mixing extracted, then be added derivatization reagent perform the derivatization reaction to get；The extractant is dichloromethane
Alkane.
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| CN201811107004.4A CN109212067B (en) | 2018-09-21 | 2018-09-21 | Detection method of high-grade fatty acid in tobacco and tobacco products and detection sample processing method |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101131378A (en) * | 2007-09-29 | 2008-02-27 | 中国烟草总公司郑州烟草研究院 | Method for detecting volatile and semi-volatile organic acid in tobacco leaf or cut tobacco |
-
2018
- 2018-09-21 CN CN201811107004.4A patent/CN109212067B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101131378A (en) * | 2007-09-29 | 2008-02-27 | 中国烟草总公司郑州烟草研究院 | Method for detecting volatile and semi-volatile organic acid in tobacco leaf or cut tobacco |
Non-Patent Citations (4)
| Title |
|---|
| 于治国 等: "《药物分析》", 31 August 2017, 中国医药科技出版社 * |
| 国家烟草专卖局: "《YC/T 470-2013 烟草及烟草制品 西柏烷二萜醇的测定气相色谱-质谱联用法》", 30 June 2013, 中国标准出版社 * |
| 葛少林 等: "烟草中挥发性与半挥发性有机酸的GC-MS法快速检测", 《香料香精化妆品》 * |
| 陈飞钦 等: "超声波提取-气相色谱/质谱联用分析烟叶中挥发性和半挥发性有机酸", 《分析科学学报》 * |
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