CN107490642A - Method that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance - Google Patents

Method that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance Download PDF

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CN107490642A
CN107490642A CN201710730193.XA CN201710730193A CN107490642A CN 107490642 A CN107490642 A CN 107490642A CN 201710730193 A CN201710730193 A CN 201710730193A CN 107490642 A CN107490642 A CN 107490642A
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alkaloids
alkaloid
cigarette
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韩书磊
陈欢
付亚宁
刘彤
李翔宇
张洪非
罗彦波
庞永强
韩雪
侯宏卫
胡清源
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National Tobacco Quality Supervision and Inspection Center
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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
    • 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
    • G01N30/62—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
    • 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
    • G01N2030/065—Preparation using different phases to separate parts of sample
    • 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

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Abstract

The present invention relates to method that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance, belong to physico-chemical analysis technical field.This method comprises the following steps：1）Using linear pattern or rotating disk smoking machine, cigarette mainstream smoke condensate is trapped with cambridge filter；2）Using 5% sodium hydrate aqueous solution by the alkaloid separate out in cambridge filter；3）Liquid liquid oscillation extraction is carried out using methylene chloride/methanol mixed solution as extractant；4）After being stored at room temperature, remove layer organic phase and progress GC MS analyses in chromatography bottle are transferred to after 0.22 μm of organic phase filter membrane filters.In the present invention 12 grow tobacco alkaloid average recovery of standard addition between 94% ~ 108%, precision is not higher than 6.8%, and method detection limit has the advantages that easy to operate, flux is high, high sensitivity, the rate of recovery and reproducible in 0.18 ~ 1.76 μ g/ branch.

Description

Method that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance
Technical field
The invention belongs to physico-chemical analysis technical field, and in particular to a kind of to determine 12 kinds of lifes in cigarette smoke granule phase substance simultaneously The method of alkaloids.
Background technology
Alkaloid refers to a kind of organic compound containing azacyclo- being present in organism（Protein, titanium class, amino Except acid, ammonia, amino sugar, nucleotides, nucleic acid）, it is general that there is alkalescence and show certain physiologically active, the cigarette of document report Careless alkaloid mainly includes nicotine, nornicotine, anabasine, myosmine, anatabine etc., wherein, nicotine content in tobacco It is very high, it is most important alkaloid in tobacco.During cigarette smoking, nicotiana alkaloids can be by alkaloid warm in cigarette shreds Crack or be directly transferred in cigarette smoke, these nicotiana alkaloids have important shadow to chemical property, the pipe pipe feature of flue gas Ring.For example, nicotine content height can typically have positive effect to suction quality, nornicotine content height typically has negatively to suction quality Effect, the alkaline breath of cigarette feature is from nicotine, nornicotine etc. caused by tobacco cracking, and wheat caused by nornicotine cracking This bright and substituted pyridine compound can then produce niff.In addition to suction quality, secondary alkaloid can also be to tobacco-induced addiction Property has an impact, and nicotine is as most important pharmaceutical compositions in cigarette smoke, the main reason for being tobacco-induced addiction, still, cigarette Secondary alkaloid outside alkali can also have certain addiction feature enhancement effect to nicotine.Therefore, in Accurate Determining cigarette smoke Alkaloid, it is significant.At present, the method for determining nicotiana alkaloids in cigarette smoke granule phase substance mainly has gas phase Chromatography, liquid chromatography, gas chromatography-mass spectrometry and with the chromatography of ions etc..But the secondary biology of these researchs Alkali is concentrated mainly on nornicotine, anatabine, anabasine, myosmine etc., and the research to other secondary alkaloids is less.
The content of the invention
The purpose of the present invention is based on above-mentioned technical deficiency, establishes a kind of while determines 12 in cigarette smoke granule phase substance The method of kind alkaloid.This method can determine 12 kinds of alkaloids in object simultaneously by a sample pretreatment process, Have the advantages that easy to operate, flux is high, high sensitivity, the rate of recovery and reproducible.
The purpose of the present invention is achieved through the following technical solutions：
Method that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance, 12 kinds of alkaloids are respectively：Nicotine, N- Methylanabasine, nornicotine, myosmine, anabasine, β-nicotyrine, anatabine, 2,3 '-bipyridyl, β-diene drop Nicotine, cotinine, harmine, nicotelline, it is characterised in that：Using linear pattern or rotating disk smoking machine, mould is aspirated by ISO, HCI Formula smoking cigarette, cigarette mainstream smoke condensate is trapped with cambridge filter, using 5% sodium hydroxide solution by cambridge filter Alkaloid separate out, then with methylene chloride/methanol be VDichloromethane/VMethanol=4:1 mixed solution carries out liquid-liquid extraction, with gas Phase chromatograph-mass spectrometer coupling method is measured, inner mark method ration；Specifically include following steps：
1）The preparation of standard working solution
Nicotine standard working solution with 5 grades of concentration gradients is prepared respectively and other 11 kinds with 12 grades of concentration gradients secondary Alkaloid standard working solution；Wherein, the quantitative of nicotine uses nornicotine -2 using quinoline as internal standard, the quantitative of nornicotine, 4,5,6-d4As internal standard, the quantitative of other secondary alkaloids uses 2,2 '-bipyridyl-d8As internal standard；
2）Sample pre-treatments
The cambridge filter that has cigarette smoke granule phase substance will be trapped to be put into 50 mL conical flasks, then to be separately added into 100 μ L internal standards molten The NaOH aqueous solution of liquid and 5.0 mL5%, after concussion mixes, stand about 10 min；Then 20.0 mL methylene chloride/methanols are added Mixed solution, tool plug oscillation extraction 40 min；After being stored at room temperature 1 h, remove layer organic phase and filtered through 0.22 μm of organic phase filter membrane After be transferred in chromatography bottle progress GC-MS analyses；
3）Gas chromatography-mass spectrometry analysis
Mass detector is equipped with using gas chromatograph to analyze made testing sample solution and standard working solution, is obtained Related chromatogram；
4）Specification Curve of Increasing and result calculate.
In the present invention, the preparation method of standard working solution is specific as follows：
The preparation of 1.1 inner mark solutions
About 5.0 g quinoline, 100.0 mg nornicotines -2,4,5,6-d are accurately weighed respectively4, 100.0 2,2 '-bipyridyls of mg- d8In 50 mL brown volumetric flasks, with methanol constant volume, internal standard storing solution is produced.Quinoline is as nicotine internal standard, nornicotine -2,4, 5,6-d4As nornicotine internal standard, and 2,2 '-bipyridyl-d8As other 10 kinds secondary alkaloid internal standards.By internal standard storing solution with 5 times of methanol dilution, that is, obtain inner mark solution.
The preparation of 1.2 primary standard storing solutions
1.2.1 the preparation of one-level nicotine standard storing solution
About 1000.0 mg nicotine accurately are weighed, is placed in 10 mL brown volumetric flask, is released with methanol and be settled to scale.The solution It should be kept in dark place under the conditions of 4 DEG C~8 DEG C.
1.2.2 the preparation of the secondary alkaloid standard reserving solution of 11 kinds of one-level
The about 20.0 secondary alkaloids of mg are accurately weighed respectively, are placed in 10 mL brown volumetric flask, are settled to methanol dilution Scale.The solution should be kept in dark place under the conditions of 4 DEG C~8 DEG C.
The preparation of 1.3 secondary standard storing solutions
1.3.1 the preparation of two level nicotine standard storing solution
About 1.0 mL one-level nicotine standard storing solutions accurately are pipetted, are placed in 10 mL brown volumetric flask, with methanol dilution constant volume To scale.The solution should be kept in dark place under the conditions of 4 DEG C~8 DEG C.
1.3.2 the preparation of the secondary alkaloid standard reserving solution of 11 kinds of two level
The secondary alkaloid standard reserving solution of 11 kinds of about 0.1 mL one-levels accurately is pipetted, is placed in 10 mL brown volumetric flask, uses first Alcohol dilution is settled to scale.The solution should be kept in dark place under the conditions of 4 DEG C~8 DEG C.
The preparation of 1.4 standard working solutions
1.4.1 the preparation of nicotine standard working solution
20 μ L, 40 μ L, 100 μ L, 200 μ L, 400 μ L two level nicotine standard storing solution are accurately pipetted respectively in different In 10 mL brown volumetric flasks, then 50 μ L inner mark solutions (quinoline) of accurate addition respectively, scale is settled to dchloromethane, Obtain the series standard solution of 5 various concentrations.
1.4.2 the preparation of 11 kinds of secondary alkaloid standard working solutions
10 μ L, 20 μ L, 50 μ L, 100 μ L, 200 μ L, 500 μ the L secondary alkaloid mark of 11 kinds of two level are accurately pipetted respectively Quasi- storing solution and 10 μ L, 20 μ L, 50 μ L, 100 μ L, 200 μ L, the 500 μ L secondary alkaloid standard inventory of 11 kinds of one-level Liquid is in 10 different mL brown volumetric flasks, then 50 μ L inner mark solutions of accurate addition respectively（Nornicotine -2,4,5,6-d4With 2, 2 '-bipyridyl-d8）, scale is settled to dchloromethane, that is, obtains the series standard solution of 12 various concentrations.
The GC-MS instrumental conditions are：
Chromatographic column：DB-35MS capillary chromatographic columns, stationary phase：（35%- phenyl）- methyl polysiloxane, specification：30 m × 0.25 mm × 0.25 μm。
Injector temperature：250 ℃；Sample size：1 μL；Carrier gas：Helium（Purity >=99.999%）, constant current mode, flow velocity： 1.0 mL/min。
Using two pin sample introductions, run respectively according to two groups of heating schedules：
Nicotine post heating schedule：Split sampling（Split ratio：100:1）；Solvent delay：5 min；Heating schedule：Initial temperature 80 DEG C, 1 min is kept, with 20 DEG C/min speed to 200 DEG C, then with 40 DEG C/min speed to 250 DEG C, keeps 5 min. Operation total time is 13.25 min.
Secondary alkaloid alkali post heating schedule：Split sampling（Split ratio：10:1）；Solvent delay：12 min；Heat up journey Sequence：80 DEG C of initial temperature, 1 min is kept, with 8 DEG C/min speed to 250 DEG C, keep 1 min, then with 40 DEG C/min's Speed keeps 5 min to 280 DEG C.Operation total time is 29 min.
Ionization mode：Electron bombardment ionization source（EI）；Ionizing energy：70 eV；Transmission line temperature：280 ℃；Ion source temperature： 250 ℃；Scan ion range：80~250 amu；Scanning of the mass spectrum mode：Select ion surveillance style（SIM）Scanning, object And interior target retention time, it is quantitative and it is qualitative selection ion parameters it is as shown in table 1.
The process of sample pre-treatments is specific as follows：
Five kinds of n-hexane, methyl tertiary butyl ether(MTBE), ethyl acetate, dichloromethane, chloroform solvents are selected to carry out extraction efficiency Compare.As a result find, n-hexane is all poor to the extraction efficiency of most of secondary alkaloid, and methyl tertiary butyl ether(MTBE) is to β-diene cigarette The effect of extracting of alkali and cotinine is very poor, and ethyl acetate is poor to the effect of extracting of cotinine, and when chloroform makees extractant, The extraction efficiency extreme difference of nornicotine, anabasine and anatabine, it may be possible to which target compound is decomposed, dichloromethane pair The synthesis extraction efficiency highest of all target compounds（Fig. 4）.According to literature survey, addition methanol can improve in dichloromethane Dichloromethane to the extraction efficiency of alkaloid, so, research compares dichloromethane and extractant pair during methanol different proportion The extraction efficiency of alkaloid（VDichloromethane/VMethanol=1:0、5:1、4:1、3:1、2:1）, as a result find, when dichloromethane and methanol volume Than for 4:When 1, to the synthesis extraction efficiency highest of alkaloid（Fig. 5）, therefore select VDichloromethane/VMethanol=4:1 is molten for final extraction Agent.
Alkaloid in tobacco needs to add a certain amount of basic solvent, and the alkaloid of wherein reference state can dissociate Out.Be separately added into experiment 2 mL0.5%, 1%, 2%, 5%, the NaOH aqueous solution of 10% 4 kind of concentration is compared, the results showed that The synthesis extraction efficiency highest of the 5%NaOH aqueous solution, avoids the relatively low extraction efficiency of relatively low NaOH concentration, turn avoid height The emulsion that NaOH concentration may be brought.
Therefore, it is ultimately determined to by the optimization of pre-treating method, Pretreatment：
Target cigarette is balanced into 48 h under conditions of temperature (22 ± 2) DEG C and relative humidity (60 ± 5) %, using linear pattern or Rotating disk smoking machine, according to the methods of GB/T 19609-2004, using the suction mode smoking cigarette such as ISO, HCI, filtered with 1 Cambridge Piece traps the total particulate matter in mainstream smoke of 2 cigarette, and after the completion of suction, the cambridge filter that trapping has cigarette smoke granule phase substance is put into In 50 mL conical flasks, then 100 μ L inner mark solutions and the 5.0 mL5% NaOH aqueous solution are separately added into, after concussion mixes, stood About 10 min.Then 20.0 mL methylene chloride/methanol mixed solutions are added（VDichloromethane/VMethanol=4:1）, tool plug oscillation extraction 40 min.After being stored at room temperature 1 h, remove layer organic phase and be transferred to after 0.22 μm of organic phase filter membrane filters in chromatography bottle and carry out GC-MS is analyzed.
Recovery of standard addition and precision
Respectively according to basic, normal, high 3 kinds it is horizontal add 12 kinds of alkaloid standard items, each pitch-based sphere replication 5 times, experiment It the results are shown in Table 2.As seen from table, for the average recovery of standard addition of 12 kinds of alkaloids between 94.80% ~ 108.67%, precision is not high In 6.8%.
The range of linearity, detection limit and quantitative limit
This research is quantified using internal standard method, using the concentration of each target compound as abscissa, the peak of analyte and internal standard compound Area ratio is that ordinate establishes standard curve.With optimal conditions, it is contemplated that the concentration range of target compound, it is determined that method The range of linearity, and corresponding concentration is as detection limit when being 3 according to object signal to noise ratio, as shown in table 3.As seen from table, mark The coefficient correlation of directrix curve is not less than 0.997, and method detection limit is in 0.18 ~ 1.76 μ g/ branch.
In a word, GC-MS high fluxs detection side that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance is established The Pretreatments such as extractant are optimized for method, this method, and instrument parameter is also optimized and determined, such as color Compose separation condition, Mass Spectrometry Conditions（Quantitative and qualitative ion etc.）, after this method optimization, by a sample pretreatment process Determine 12 kinds of alkaloids in cigarette smoke simultaneously, have pretreatment process is easy to operate, analysis method flux is high, the rate of recovery and The advantages that reproducible.
Brief description of the drawings
Fig. 1 is that nicotine standard working solution selects ion flow graph;
Fig. 2 is that 11 kinds of secondary alkaloid standard working solutions select ion flow graph,
In Fig. 2：1：N- methylanabasines；2：Nornicotine and nornicotine -2,4,5,6-d4；3：Myosmine；4：2,2 '-connection pyrrole Pyridine-d8；5：Anabasine；6：β-nicotyrine；7：Anatabine；8：2,3 '-bipyridyl；9：β-diene nornicotine；10：It can replace Rather；11：Harmine；12：Nicotelline.
Fig. 3 is 12 kinds of alkaloids and its structural formula
Fig. 4 is extraction efficiency schematic diagram of the different extractants to target compound（Through normalization）.
Fig. 5 is to be added in dichloromethane after different proportion methanol to the extraction efficiency schematic diagram of target compound（Through normalizing Change）.
Embodiment
The present invention is further described by specific examples below, but does not limit the present invention.
Embodiment 1：
1st, instrument and reagent consumptive material
Instrument：Gas chromatograph-mass spectrometer (GC-MS)（U.S.'s Agilent 7890B-5977A types）；SM450 linear pattern smoking machine (English Si Rulin companies of state)；AE163 electronic balances（Sensibility reciprocal：0.0001 g, Mettler companies of Switzerland）；HY-8 velocity-modulated oscillators are (often Zhou Guohua Electrical Appliances Co., Ltd.
Reagent consumptive material：Nornicotine is bought by alfa, and other 11 alkaloids that grow tobacco are bought by TRC, nornicotine -2,4, and 5, 6-d4（CDN Isotopes）, 2,2 '-bipyridyl-d8（CDN Isotopes）, methanol（DUKSAN, chromatographically pure）, dichloromethane （DUKSAN, chromatographically pure）, sodium hydroxide（Traditional Chinese medicines, analysis are pure）, water used is by Milli-Q systems（Milford, MA, USA）System ；The mm cambridge filters of Φ 44（German Borgwaldt companies）.
2nd, sample pre-treatments
Target cigarette is balanced into 48 h under conditions of temperature (22 ± 2) DEG C and relative humidity (60 ± 5) %, inhaled using linear pattern Cigarette machine, according to the methods of GB/T 19609-2004, using HCI suction mode smoking cigarettes, 2 volumes are trapped with 1 cambridge filter The total particulate matter in mainstream smoke of cigarette, after the completion of suction, the cambridge filter that trapping has cigarette smoke granule phase substance is put into 50 mL conical flasks In, then 100 μ L inner mark solutions and the 5.0 mL5% NaOH aqueous solution are separately added into, after concussion mixes, stand about 10 min.So After add 20.0 mL methylene chloride/methanol mixed solutions（VDichloromethane/VMethanol=4:1）, tool plug oscillation extraction 40 min.It is stored at room temperature After 1 h, remove layer organic phase and progress GC-MS analyses in chromatography bottle are transferred to after 0.22 μm of organic phase filter membrane filters.
3rd, instrumental conditions
Chromatographic column：DB-35MS capillary chromatographic columns, stationary phase：（35%- phenyl）- methyl polysiloxane, specification：30 m × 0.25 mm × 0.25 μm。
Injector temperature：250 ℃；Sample size：1 μL；Carrier gas：Helium（Purity >=99.999%）, constant current mode, flow velocity： 1.0 mL/min。
Using two pin sample introductions, run respectively according to two groups of heating schedules：
Nicotine post heating schedule：Split sampling（Split ratio：100:1）；Solvent delay：5 min；Heating schedule：Initial temperature 80 DEG C, 1 min is kept, with 20 DEG C/min speed to 200 DEG C, then with 40 DEG C/min speed to 250 DEG C, keeps 5 min. Operation total time is 13.25 min.
Secondary alkaloid alkali post heating schedule：Split sampling（Split ratio：10:1）；Solvent delay：12 min；Heat up journey Sequence：80 DEG C of initial temperature, 1 min is kept, with 8 DEG C/min speed to 250 DEG C, keep 1 min, then with 40 DEG C/min's Speed keeps 5 min to 280 DEG C.Operation total time is 29 min.
Ionization mode：Electron bombardment ionization source（EI）；Ionizing energy：70 eV；Transmission line temperature：280 ℃；Ion source temperature： 250 ℃；Scan ion range：80~250 amu；Scanning of the mass spectrum mode：Select ion surveillance style（SIM）Scanning, object And interior target retention time, it is quantitative and it is qualitative selection ion parameters it is as shown in table 1.
According to said determination method, in Canadian depth suction mode（HCI）Under, measure in a kind of cigarette smoke granule phase substance 12 kinds of alkaloids are as shown in the table（Unit：Nicotine is mg/ branch, and related alkaloids are μ g/ branch）：
Nicotine Nornicotine N- methylanabasines Myosmine Anabasine β-nicotyrine Anatabine 2,3 '-bipyridyl β-diene nornicotine Cotinine Harmine Nicotelline
1.10 0.35 4.92 7.40 1.32 3.33 4.08 4.27 2.49 8.48 1.64 3.64

Claims (1)

1. method that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance, 12 kinds of alkaloids are respectively：Nicotine, N- methylanabasines, nornicotine, myosmine, anabasine, β-nicotyrine, anatabine, 2,3 '-bipyridyl, β-diene Nornicotine, cotinine, harmine, nicotelline, it is characterised in that：Using linear pattern or rotating disk smoking machine, aspirated by ISO, HCI Pattern smoking cigarette, cigarette mainstream smoke condensate is trapped with cambridge filter, using 5% sodium hydroxide solution by cambridge filter In alkaloid separate out, then with methylene chloride/methanol be VDichloromethane/VMethanol=4:1 mixed solution carries out liquid-liquid extraction, with Gas chromatography-mass spectrometry is measured, inner mark method ration；Specifically include following steps：
1）The preparation of standard working solution
Nicotine standard working solution with 5 grades of concentration gradients is prepared respectively and other 11 kinds with 12 grades of concentration gradients secondary Alkaloid standard working solution；Wherein, the quantitative of nicotine uses nornicotine -2 using quinoline as internal standard, the quantitative of nornicotine, 4,5,6-d4As internal standard, the quantitative of other secondary alkaloids uses 2,2 '-bipyridyl-d8As internal standard；
2）Sample pre-treatments
The cambridge filter that has cigarette smoke granule phase substance will be trapped to be put into 50 mL conical flasks, then to be separately added into 100 μ L internal standards molten The NaOH aqueous solution of liquid and 5.0 mL5%, after concussion mixes, stand about 10 min；Then 20.0 mL methylene chloride/methanols are added Mixed solution, tool plug oscillation extraction 40 min；After being stored at room temperature 1 h, remove layer organic phase and filtered through 0.22 μm of organic phase filter membrane After be transferred in chromatography bottle progress GC-MS analyses；
3）Gas chromatography-mass spectrometry analysis
Mass detector is equipped with using gas chromatograph to analyze made testing sample solution and standard working solution, is obtained Related chromatogram, instrumental conditions when GC-MS is analyzed are：
Chromatographic column：DB-35MS capillary chromatographic columns, stationary phase：（35%- phenyl）- methyl polysiloxane, specification：30 m × 0.25 mm × 0.25 μm；
Injector temperature：250 ℃；Sample size：1 μL；Carrier gas：Helium, constant current mode, flow velocity：1.0 mL/min；
Using two pin sample introductions, run respectively according to two groups of heating schedules：
Nicotine post heating schedule：Split sampling, split ratio：100:1；Solvent delay：5 min；Heating schedule：Initial temperature 80 DEG C, 1 min is kept, with 20 DEG C/min speed to 200 DEG C, then with 40 DEG C/min speed to 250 DEG C, keeps 5 min； Operation total time is 13.25 min；
Secondary alkaloid alkali post heating schedule：Split sampling, split ratio：10:1；Solvent delay：12 min；Heating schedule：Initially 80 DEG C of temperature, 1 min is kept, with 8 DEG C/min speed to 250 DEG C, keep 1 min, then with 40 DEG C/min speed extremely 280 DEG C, keep 5 min；Operation total time is 29 min；
Ionization mode：Electron bombardment ionization source（EI）；Ionizing energy：70 eV；Transmission line temperature：280 ℃；Ion source temperature：250 ℃；Scan ion range：80~250 amu；Scanning of the mass spectrum mode：Select ion surveillance style（SIM）Scanning, object and interior Target retention time, it is quantitative and it is qualitative selection ion parameters it is as shown in the table；
4）Specification Curve of Increasing and result calculate.
CN201710730193.XA 2017-08-23 2017-08-23 Method that is a kind of while determining 12 kinds of alkaloids in cigarette smoke granule phase substance Pending CN107490642A (en)

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Cited By (3)

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CN109725098A (en) * 2019-03-04 2019-05-07 陕西中烟工业有限责任公司 A kind of flue gas rhynchophyllin rate of transform and filter tip rhynchophyllin rejection measuring method
CN110823650A (en) * 2019-10-21 2020-02-21 河南中烟工业有限责任公司 Method for collecting and detecting alkaloid in cigarette mainstream smoke
CN113933430A (en) * 2021-10-22 2022-01-14 中国烟草总公司郑州烟草研究院 High-sensitivity and high-precision analysis method suitable for detecting organic sulfur compounds in cigarette mainstream smoke

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Application publication date: 20171219