CN104090043A - Method for measuring methyleugenol in cigarette mainstream smoke - Google Patents

Abstract

The invention discloses a method for measuring methyleugenol in cigarette mainstream smoke. The method comprises the following steps: preparing an internal standard solution, a standard working solution and a sample solution, performing gas chromatography-tandem mass spectrometry analysis and calculating the measurement result. 3,4-methylene dioxo acetophenone serves as an internal standard substance for quantifying, the optimized detection method is simple and convenient in operation, sensitive in response and accurate in quantitative analysis, interference brought by a complex sample matrix is effectively reduced, the target has good signal response and has high linear correlation due to the adopted chromatography-mass spectrum conditions, the detection limit is 0.31ng per cigarette, the relative standard deviation is 4.30 percent, and the adding standard recovery is between 94.85 percent and 112.51 percent. Therefore, the method is high in sensitivity, high in repeatability and high in recovery rate and is suitable for quantitative analysis of trace targets in the complex smoke matrix.

Description

A kind of method of measuring methyl eugenol in cigarette mainstream flue gas

Technical field

The invention belongs to cigarette physical and chemical index detection technique field, be specifically related to a kind of method of measuring methyl eugenol in cigarette mainstream flue gas.

Background technology

Methyl eugenol, have another name called eugenol methyl ether, left-handed rose oxide, eugenol methyl ether, O-methyl eugenol, 4-allyl verasine, 4-allyl veratrole, eugenol ylmethyl ether, eugenyl methyl ether, lure fly ether, English Methyl Eugenol (Eugenyl Methyl Ether) by name.Be for No. CAS 93-15-2, molecular formula is C ₁₁h ₁₄o ₂, molecular weight is 178.23, colourless to micro-yellow liquid.249 DEG C of boiling points, relative density 1.032-1.036, refractive index 1.532-1.536,99 DEG C of flash-points, are dissolved in ethanol and oils.Have the pungent fragrance of fresh and sweet cloves-fennel, like carnation breath, fragrance is lasting compared with thoroughly sending out, and has the pungent fragrance of gentleness of tea sample.According to toxic grade classification, methyl eugenol is poisoning.According to MSDS data information, its LD50 (rat, per os) is 1179 mgs/kg, and LD50 (mouse, abdominal cavity) is 540 mgs/kg.But the potential hazard of methyl eugenol is familiar with by domestic and international, methyl eugenol has been listed in the forbidding name list of China's " cosmetics health specification ".

For the analyzing and testing of methyl eugenol, abroad, the identified and quantitative analysis in some spice berries of alkylene benzene class aromatic substance.In the time of the alkylene benzene class aromatic substance of analyzing in plant, the instrument analytical method adopting mainly contains GC or GC-MS, and HPLC.Such as 1985, the people such as Carman A S used GC method to carry out fast quantitative analysis to methyl eugenol, myristicin in fresh and frozen carrot; 1988, Archer A W used HPLC method to carry out quantitative analysis to the methyl eugenol in nutmeg and nutmeg dry hide, myristicin; In the time of the alkylene benzene class aromatic substance of analyzing in plant, the Sample Pretreatment Technique adopting, according to the difference of sample substrate, mainly contain solvent extraction (Schmelta, 1967), Solid-Phase Extraction (Yates and England, nineteen eighty-two), steam distillation (Lavoie et al, 1985), supercritical fluid extraction (Heikes, 1994), solid-phase microextraction (Clark and Bunch, 1997).In addition, the report that also has the alkylene benzene class aromatic substance in pair tobacco to analyze, such as 1999, Stanfill and Ashley used the alkylene benzene class aromatic substance in SPME-GC-MS method contrast enlightening cigarette and cloves cigarette to carry out quantitative test.But for cigarette smoke, due to complicacy and the lower reason of object content of flue gas matrix, the quantitative test of alkylene benzene class aromatic substance is considered to be worth the problem of challenge always.1964, Rodgman and Cook collected 20560 granule phase substances without filter tip cigarette flue gas, adopt suitable pre-treating method, separated and identified eugenol, isoeugenol and methyl eugenol.1966, Schmelta collected 1kg cigarette smoke granule phase substance (being equivalent to the product of 50000 cigarette burnings), analyzed myristicin wherein.1986, the people such as Lavoie et al used GC-MS method to carry out analyzing to the eugenol in cloves cigarette and measure.Until 2000, the people such as Stephen adopt Solid-Phase Extraction-GC-MS method, the cigarette of selling on American market is comprised to 8 kinds of alkylene benzene class aromatic substances such as eugenol, methyl eugenol analyze mensuration, and analyze the impact of different filter tip degree of ventilation on alkylene benzene class aromatic substance burst size.Wherein, in the time of sample pre-treatments, according to the different content of object, be divided into two parts, i.e. only solid-phase extraction column (Pre-SPE) and mistake solid-phase extraction column (Post-SPE) of extract.

At home, some researchers are analyzed and researched to the alkylene benzene class aromatic substance in plant, cosmetics and essence and flavoring agent.Such as, 1997, the people such as Wang Dong carried out quantitative test to the contained methyl eugenol of Herba Asari different growing stages and safrole by vapor-phase chromatography.1999, the people such as Zhou Changzheng adopt internal standard method to 12 kinds of the Domestic Drugs root of Chinese wild ginger in totally 24 sample volatile oil the content of methyl eugenol and safrole carried out gas chromatographic analysis.2009, the people such as Wu Ting set up the different gas chromatographies that add in Fragrant food taking Isosorbide-5-Nitrae-dibromobenzene and phenethyl acetate as fixed natural nuisance absinthol, esdragol and methyl eugenol content of interior mapping, and retention time is qualitative, inner mark method ration.2012, the people such as Chen Jianwei adopted supercritical fluid extraction respectively Herba Asari root and rhizome to be extracted and separated with steam distillation, obtained respectively 5.5%SFE extract and 2.8% volatile oil.Application GC-MS analyzes, and identifies 7 kinds of chemical compositions from Herba Asari SFE extract, accounts for 72.70% of extract total amount, and wherein methyl eugenol is 44.62%; From Herba Asari volatile oil, identify 19 kinds of chemical compositions, account for 88.53% of volatile oil total amount, wherein methyl eugenol is 43.02%.Two kinds of methods have 6 kinds of compositions identical, and wherein carcinogen safrole SFE method is lower 2.8 times than the way of distillation.2012, the people such as Zhu Xiaolan have set up ultrasonic solvent extracts and dispersive liquid-liquid microextraction (DLLME) combines extraction, have purified pretreatment technology, employing GC-MS Analysis 8 kinds of alkyl benzene fragrance nuisances such as the absinthol in cigarette additive, esdragol, pulegone, methyl eugenol, cumarin, 6-Methylcoumarin and ayapanin.The beautiful analytical approach that waits people to set up methyl eugenol in Gas Chromatography-Mass Spectrometry perfume of phase.

In sum, mainly adopt the method such as GC-MS, HPLC to analyze to the detection of the alkylene benzene class aromatic substances such as methyl eugenol both at home and abroad.According to the difference of sample substrate, pre-treating method used mainly contains solvent extraction, Solid-Phase Extraction, steam distillation, supercritical fluid extraction, solid-phase microextraction and while distillation extraction.

The sample that above analyzing and testing relates to is mainly spice berry, lack the analysis and research to methyl eugenol in cigarette smoke, only report is that Stephen in 2000 and Ashley adopt Solid-Phase Extraction-GC/MS method, and methyl eugenol in the cigarette smoke of selling on American market is analyzed to mensuration at 8 kinds of interior alkylene benzene class aromatic substances.But based on the repeated authentication to the method, on the solid-phase extraction column of the CN bonding that discovery methyl eugenol adopts at document, retention is not strong, does not reach the object of selective elution, at all cannot accurate quantitative analysis evaluating objects thing.

Summary of the invention

Goal of the invention: for the deficiencies in the prior art, the object of this invention is to provide a kind of method of measuring methyl eugenol in cigarette mainstream flue gas, to fast, accurately detecting the content of methyl eugenol in cigarette mainstream flue gas, measurement result is accurate, matrix interference is few.

Technical scheme: in order to realize foregoing invention object, the technical solution used in the present invention is as follows:

A method of measuring methyl eugenol in cigarette mainstream flue gas, comprises the following steps:

(1) preparation of inner mark solution: for internal standard compound, make solvent with ethanol with 3,4-(methylenedioxy) acetophenone, prepare inner mark solution;

(2) preparation of standard operation solution: taking methyl eugenol as standard items, make solvent with ethanol, be prepared into standard inventory solution through stepwise dilution, then add respectively a certain amount of inner mark solution, be prepared into standard operation solution;

(3) preparation of sample solution: collected the object in cigarette smoke, accurately added a certain amount of inner mark solution and extraction solvent, mechanical shaking extraction; Extraction parts extract, through Solid-Phase Extraction column purification, after scavenging solution is concentrated, obtains sample solution;

(4) gas chromatography-Tandem Mass Spectrometry Analysis: standard operation solution and sample solution are detected to analysis with gas chromatography-tandem mass spectrometer (GC-MS/MS); Chromatographiccondition is: adopt HP-5MS (30m × 0.25mm × 0.25 _μm) capillary chromatographic column; Carrier gas is helium, and constant current speed is 1mL/min; Sample size is 1 μ L, adopts pulse Splitless injecting samples; Injector temperature is 250 DEG C; Transmission line temperature is 280 DEG C; Heating schedule is that initial temperature is 60 DEG C, is increased to 160 DEG C, then is increased to 300 DEG C with the speed of 10 DEG C/min with the speed of 4 DEG C/min; Mass spectrophotometry condition is: ionization mode is EI source, positive ion mode; Ion source temperature: 230 DEG C; Quadrupole rod temperature: be 150 DEG C; Collision gas: nitrogen, flow velocity 1.5mL/min, carrier gas (helium) flow velocity is 2.25mL/min; Multiple-reaction monitoring (MRM) pattern, detail parameters is listed in table 1;

The MRM parameter of table 1 object

(5) drafting of standard working curve and the calculating of sample result: taking methyl eugenol in standard operation solution with the ratio of interior target concentration as horizontal ordinate, taking methyl eugenol in chromatogram with the ratio of interior target peak area as ordinate, carry out linear regression analysis, obtain standard working curve.By methyl eugenol in the sample solution recording under the same terms and interior target chromatographic peak area ratio, substitution standard working curve, converts and tries to achieve the content of methyl eugenol in cigarette sample main flume.

In step (1), the preparation of described inner mark solution specifically comprises the following steps: mark storing solution 1): accurately take 100mg internal standard compound 3,4-(methylenedioxy) acetophenone, with ethanol constant volume, to 100mL, obtaining concentration is the interior mark storing solution of 1000 μ g/mL; 2) inner mark solution: accurately pipette mark storing solution in 1mL, to 100mL, obtaining concentration is the interior mark storing solution of 10 μ g/mL with ethanol constant volume.

In step (2), the preparation of described standard operation solution specifically comprises the following steps: 1) primary standard storing solution: accurately take 100mg methyl eugenol, with ethanol constant volume, to 100mL, obtaining concentration is the primary standard storing solution of 1000 μ g/mL; 2) secondary standard storing solution: accurately pipette 0.5mL primary standard storing solution, to 100mL, obtaining concentration is 5 with ethanol constant volume _μthe secondary standard storing solution of g/mL; 3) standard operation solution: accurately pipette respectively secondary standard storing solution 10 μ L, 20 μ L, 50 μ L, 100 μ L and 200 μ L, and the each 100 μ L of inner mark solution, be settled to as solvent in the volumetric flask of 10mL with ethanol, obtain the series standard working solution that concentration is respectively 5ng/mL, 10ng/mL, 25ng/mL, 50ng/mL and 100ng/mL, wherein interior mark concentration is 100ng/mL.

In step (3), the preparation of described sample solution specifically comprises the following steps: 1) cigarette smoking: the TPM of collecting 20 cigarette according to the requirement of GB/T19609 standard; 2) sample extraction: the trapping that upper step is obtained has the glass fiber filter of TPM to be positioned over that in 100mL conical flask, (filter disc is answered full wafer to put into conical flask and paved, can not shred or tear up), accurately add 0.4mL inner mark solution to exist, and add 40mL normal hexane, be placed in Clothoid type oscillator oscillation extraction 40min under 160 turn/min conditions, leave standstill a moment; 3) extract purifies: accurately pipette 10mL sample extraction liquid to solid-phase extraction column (silica gel that filler is 500mg), until extract toward be displaced downwardly to extraction pillar top after, dividing adds for 2 times 10mL normal hexane to carry out drip washing, drip washing speed remains on 1～2mL/min, collects all leacheates; 4) leacheate is concentrated: the concentrated bottle that fills all leacheates is connected on Rotary Evaporators; rotary evaporation under 80 DEG C, high pure nitrogen protection, or under 55 DEG C, vacuum condition rotary evaporation, be concentrated to 1mL left and right; obtain sample feeding liquid, pending GC-MS/MS analyzes.

In step (5), the regression equation corresponding with standard working curve is Y=0.2017X-0.0012, coefficient R ²be 0.9997, detect and be limited to 0.31ng/ and prop up, concentration range is 5.25～105ng/mL.

In step (5), then, the computing formula of trying to achieve the content of methyl eugenol in cigarette sample is as follows:

$W=\frac{(\frac{A}{\mathrm{As}}-b)\×\mathrm{Ms}}{a\×M}$

In formula: W is the content of methyl eugenol in cigarette sample, unit is that ng/ props up; A is the peak area of methyl eugenol; As is interior mark 3, the peak area of 4-(methylenedioxy) acetophenone; B is the intercept of standard working curve; Ms is interior target quality in sample solution, and unit is ng; A is the slope of standard working curve; M is the quantity of cigarette, and unit is for propping up.

Beneficial effect: compared with prior art, the present invention has following excellent results:

(1) the inventive method is measured the content of methyl eugenol in cigarette mainstream flue gas first based on gas chromatography-tandem mass spectrometer, solve in flue gas methyl eugenol burst size level low, and the impact of flue gas complex matrices on the factor such as the Analysis interference of object is serious.

(2) the inventive method is utilized inner mark method ration, precisely constant volume, and can reduce the error of being brought by pre-treating method reappearance and instrument precision problem.

(3) to have selected peak shape and the best HP-5MS post of separating effect be separating column to the inventive method, and adopt the GC-MS/MS (MRM) that can simultaneously provide quantitative and qualitative analysis information as detecting pattern, the sensitivity that has improved response signal, has ensured precision of analysis.

Brief description of the drawings

Fig. 1 is the process flow diagram of assay method of the present invention;

Fig. 2 is the extraction ion pair chromatogram of methyl eugenol in standard operation solution; Wherein, upper figure is 178-> 107 ion pairs, and figure below is 178-> 147 ion pairs;

Fig. 3 is the extraction ion pair chromatogram of methyl eugenol in sample solution; Wherein, upper figure is 178-> 107 ion pairs, and figure below is 178-> 147 ion pairs;

Fig. 4 is the extraction ion pair chromatogram of internal standard compound in standard operation solution; Wherein, upper figure is 164-> 121 ion pairs, and figure below is 164-> 149 ion pairs;

Fig. 5 is the extraction ion pair chromatogram of internal standard compound in sample solution; Wherein, upper figure is 164-> 121 ion pairs, and figure below is 164-> 149 ion pairs.

Embodiment

Below in conjunction with specific embodiment, the present invention is described further.

Embodiment 1

Measure a method for methyl eugenol in cigarette mainstream flue gas, process flow diagram as shown in Figure 1, comprises the steps:

(1) preparation of inner mark solution

Interior mark storing solution: accurately take 0.1069g internal standard compound 3,4-(methylenedioxy) acetophenone, with ethanol constant volume, to 100mL, obtaining concentration is the interior mark storing solution of 1069 μ g/mL; Inner mark solution: accurately pipette mark storing solution in 1mL, to 100mL, obtaining concentration is the interior mark storing solution of 10.69 μ g/mL with ethanol constant volume.

(2) preparation of standard operation solution

Primary standard storing solution: accurately take 0.1050mg methyl eugenol, to 100mL, obtaining concentration is the primary standard storing solution of 1050 μ g/mL with ethanol constant volume; Secondary standard storing solution: accurately pipette 0.5mL primary standard storing solution, to 100mL, obtaining concentration is the secondary standard storing solution of 5.25 μ g/mL with ethanol constant volume; Standard operation solution: accurately pipette respectively secondary standard storing solution 10 μ L, 20 μ L, 50 μ L, 100 μ L and 200 μ L, and the each 100 μ L of inner mark solution, be settled in the volumetric flask of 10mL as solvent with ethanol, obtain series standard working solution.This series standard solution concentration is respectively 5.25ng/mL, 10.5ng/mL, 26.25ng/mL, 52.5ng/mL and 105ng/mL, and wherein interior mark concentration is 106.9ng/mL.

(3) preparation of sample solution

1. cigarette smoking: the TPM of collecting 20 cigarette according to the requirement of GB/T19609 standard.2. sample extraction: the trapping that upper step is obtained has the glass fiber filter of TPM to be positioned over that in 100mL conical flask, (filter disc is answered full wafer to put into conical flask and paved, can not shred or tear up), accurately add 0.4mL inner mark solution to exist, and add 40mL normal hexane, be placed in Clothoid type oscillator oscillation extraction 40min under 160 turn/min conditions, leave standstill a moment.3. extract purifies: accurately pipette 10mL sample extraction liquid to solid-phase extraction column (silica gel that filler is 500mg), until extract toward be displaced downwardly to extraction pillar top after, divide and add for 2 times 10mL normal hexane to carry out drip washing, drip washing speed remains on 1～2mL/min, collects all leacheates.4. leacheate is concentrated: the concentrated bottle that fills all leacheates is connected on Rotary Evaporators; rotary evaporation under 80 DEG C, high pure nitrogen protection (or under 55 DEG C, vacuum condition rotary evaporation); be concentrated to 1mL left and right, obtain sample feeding liquid, pending GC-MS/MS analyzes.

(4) gas chromatography-Tandem Mass Spectrometry Analysis

Standard operation solution and the sample solution to be measured of getting respectively 5 variable concentrations carry out gas chromatography-Tandem Mass Spectrometry Analysis.Its chromatographiccondition is: adopt HP-5MS (m) capillary chromatographic column of 30m × 0.25mm × 0.25 μ; Carrier gas is helium, and constant current speed is 1mL/min; Sample size is 1 μ L, adopts pulse Splitless injecting samples; Injector temperature is 250 DEG C; Transmission line temperature is 280 DEG C; Heating schedule is that initial temperature is 60 DEG C, is increased to 160 DEG C, then is increased to 300 DEG C with the speed of 10 DEG C/min with the speed of 4 DEG C/min.Its mass spectrophotometry condition is: ionization mode is EI source, positive ion mode; Ion source temperature: 230 DEG C; Quadrupole rod temperature: be 150 DEG C; Collision gas: nitrogen, flow velocity 1.5mL/min, carrier gas (helium) flow velocity is 2.25mL/min; Multiple-reaction monitoring (MRM) pattern, detail parameters is listed in table 1.The extraction ion pair chromatogram of safrole in standard operation solution, as shown in Figure 2, wherein, upper figure is 178-> 107 ion pairs, figure below is 178-> 147 ion pairs; The extraction ion pair chromatogram of safrole in sample solution, as shown in Figure 3, wherein, upper figure is 178-> 107 ion pairs, figure below is 178-> 147 ion pairs; The extraction ion pair chromatogram of internal standard compound in standard operation solution, as shown in Figure 4, wherein, upper figure is 164-> 121 ion pairs, figure below is 164-> 149 ion pairs; The extraction ion pair chromatogram of internal standard compound in sample solution, as shown in Figure 5, wherein, upper figure is 164-> 121 ion pairs, figure below is 164-> 149 ion pairs.

The MRM parameter of table 1 object

(5) Specification Curve of Increasing and result are calculated

First, taking methyl eugenol in standard operation solution with the ratio of interior target concentration as horizontal ordinate, taking methyl eugenol in chromatogram with the ratio of interior target peak area as ordinate, carry out linear regression analysis, obtain standard working curve, get the standard operation solution of least concentration, do 9 Parallel testing analyses, calculate its standard deviation, taking concentration corresponding to the standard deviations of 3 times as detection limit.The regression equation corresponding with standard working curve is Y=0.2017X-0.0012, coefficient R ²be 0.9997, detect and be limited to 0.31ng/ and prop up, concentration range is 5.25～105ng/mL.

Then, by methyl eugenol in the sample solution recording under the same terms and interior target chromatographic peak area ratio, substitution standard working curve, tries to achieve the content of methyl eugenol in cigarette sample, and computing formula is as follows:

$W=\frac{(\frac{A}{\mathrm{As}}-b)\×\mathrm{Ms}}{a\×M}$

In formula:

W---the content of methyl eugenol in cigarette sample, unit is every of nanogram (ng/ props up);

The peak area of A---methyl eugenol;

As---interior mark 3, the peak area of 4-(methylenedioxy) acetophenone;

The intercept of b---standard working curve;

Ms---interior target quality in sample solution, unit is nanogram (ng);

The slope of a---standard working curve;

The quantity of M---cigarette, unit is for propping up.

In the present embodiment, measure 5 methyl eugenol content in different brands cigarette sample, testing result sees the following form 2.

Table 2 sample detection result

Sample number into spectrum	Cigarette type	Detected value (ng/ props up)
			Cigarette A	Mixed type	2.46
Cigarette B	Mixed type	3.52
			Cigarette C	Fire-cured tobacco type	2.92
Cigarette D	Fire-cured tobacco type	5.13
			Cigarette E	Fire-cured tobacco type	3.82

Embodiment 2

Taking a cigarette sample as analytic target, adopt the method for embodiment 1 to measure the content of object, carry out Precision Experiment, measure altogether 6 times, calculate respectively the relative standard deviation (RSD) of 6 replicate determination results, measurement result sees the following form 3.In table, result shows, the repeated RSD<5% of this experimental technique.For trace materials analyzing and testing, the method has good precision.

The precision of table 3 method

Sequence number	Measurement result (ng/ props up)
		1	2.89
2	2.69
		3	2.75
4	2.76
		5	2.56
6	2.63
		RSD(％)	4.30

Taking a cigarette sample as object, carry out the mark-on processing of basic, normal, high three variable concentrations levels, then process according to above-mentioned standard pre-treatment step, carry out again GC-MS/MS analysis, the content of measuring object, calculates its recovery of standard addition, the results are shown in following table 4.Can find out from following table, the recovery of object is between 94.85%～112.51%.On the whole, for the object for content in several nanogram levels, the method has the recovery relatively preferably.

The recovery of table 4 method

The present embodiment standard solution used only describes as an example of one of them concentration example, typical curve and regression equation that the prepared standard solution of other concentration value obtains through gas chromatography-Tandem Mass Spectrometry Analysis are same as the previously described embodiments, are not enumerating at this.Just method for a better understanding of the present invention of illustrated embodiment, does not have any restriction, and said method or the method that is equal to above-mentioned situation are all included in the protection domain of technical scheme of the present invention.

Claims (6)

1. a method of measuring methyl eugenol in cigarette mainstream flue gas, is characterized in that, comprises the steps:

(1) preparation of inner mark solution: for internal standard compound, make solvent with ethanol with 3,4-(methylenedioxy) acetophenone, prepare inner mark solution;

(2) preparation of standard operation solution: taking methyl eugenol as standard items, make solvent with ethanol, be prepared into standard inventory solution through stepwise dilution, then add respectively quantitative inner mark solution, be prepared into standard operation solution;

(3) preparation of sample solution: collected the object in cigarette smoke, accurately added quantitative inner mark solution and extraction solvent, mechanical shaking extraction; Extraction parts extract, through Solid-Phase Extraction column purification, after scavenging solution is concentrated, obtains sample solution;

(4) gas chromatography-Tandem Mass Spectrometry Analysis: standard operation solution and sample solution are detected to analysis with gas chromatography-tandem mass spectrometer; Chromatographiccondition is: adopt HP-5MS capillary chromatographic column; Carrier gas is helium, and constant current speed is 1mL/min; Sample size is 1 μ L, adopts pulse Splitless injecting samples; Injector temperature is 250 DEG C; Transmission line temperature is 280 DEG C; Heating schedule is that initial temperature is 60 DEG C, is increased to 160 DEG C, then is increased to 300 DEG C with the speed of 10 DEG C/min with the speed of 4 DEG C/min; Mass spectrophotometry condition is: ionization mode is EI source, positive ion mode; Ion source temperature: 230 DEG C; Quadrupole rod temperature: be 150 DEG C; Collision gas: nitrogen, flow velocity 1.5mL/min, flow rate of carrier gas is 2.25mL/min; Multiple-reaction monitoring pattern, detail parameters is listed in table 1;

The MRM parameter of table 1 object

(5) drafting of standard working curve and the calculating of sample result: taking methyl eugenol in standard operation solution with the ratio of interior target concentration as horizontal ordinate, taking methyl eugenol in chromatogram with the ratio of interior target peak area as ordinate, carry out linear regression analysis, obtain standard working curve.By methyl eugenol in the sample solution recording under the same terms and interior target chromatographic peak area ratio, substitution standard working curve, converts and tries to achieve the content of methyl eugenol in cigarette sample main flume.

2. the method for methyl eugenol in mensuration cigarette mainstream flue gas according to claim 1, it is characterized in that: in step (1), the preparation of described inner mark solution specifically comprises the following steps: mark storing solution 1): accurately take 100mg internal standard compound 3,4-(methylenedioxy) acetophenone, with ethanol constant volume, to 100mL, obtaining concentration is the interior mark storing solution of 1000 μ g/mL; 2) inner mark solution: accurately pipette mark storing solution in 1mL, to 100mL, obtaining concentration is the interior mark storing solution of 10 μ g/mL with ethanol constant volume.

3. the method for methyl eugenol in mensuration cigarette mainstream flue gas according to claim 1, it is characterized in that: in step (2), the preparation of described standard operation solution specifically comprises the following steps: 1) primary standard storing solution: accurately take 100mg methyl eugenol, with ethanol constant volume, to 100mL, obtaining concentration is the primary standard storing solution of 1000 μ g/mL; 2) secondary standard storing solution: accurately pipette 0.5mL primary standard storing solution, to 100mL, obtaining concentration is 5 with ethanol constant volume _μthe secondary standard storing solution of g/mL; 3) standard operation solution: accurately pipette respectively secondary standard storing solution 10 μ L, 20 μ L, 50 μ L, 100 μ L and 200 μ L, and the each 100 μ L of inner mark solution, be settled to as solvent in the volumetric flask of 10mL with ethanol, obtain the series standard working solution that concentration is respectively 5ng/mL, 10ng/mL, 25ng/mL, 50ng/mL and 100ng/mL, wherein interior mark concentration is 100ng/mL.

4. the method for methyl eugenol in mensuration cigarette mainstream flue gas according to claim 1, it is characterized in that: in step (3), the preparation of described sample solution specifically comprises the following steps: 1) cigarette smoking: the TPM of collecting 20 cigarette according to the requirement of GB/T19609 standard; 2) sample extraction: the trapping that upper step is obtained has the glass fiber filter of TPM to be positioned in 100mL conical flask, accurately add 0.4mL inner mark solution to exist, and add 40mL normal hexane, and be placed in Clothoid type oscillator oscillation extraction 40min under 160 turn/min conditions, leave standstill a moment; 3) extract purifies: accurately pipette 10mL sample extraction liquid to solid-phase extraction column,, divide and add for 2 times 10mL normal hexane to carry out drip washing toward being displaced downwardly to behind the top of extraction pillar until extract, drip washing speed remains on 1～2mL/min, collects all leacheates; 4) leacheate is concentrated: the concentrated bottle that fills all leacheates is connected on Rotary Evaporators; rotary evaporation under 80 DEG C, high pure nitrogen protection, or under 55 DEG C, vacuum condition rotary evaporation, be concentrated to 1mL left and right; obtain sample feeding liquid, pending GC-MS/MS analyzes.

5. the method for methyl eugenol in mensuration cigarette mainstream flue gas according to claim 1, is characterized in that: in step (5), the regression equation corresponding with standard working curve is Y=0.2017X-0.0012, coefficient R ²be 0.9997, detect and be limited to 0.31ng/ and prop up, concentration range is 5.25～105ng/mL.

6. the method for methyl eugenol in mensuration cigarette mainstream flue gas according to claim 1, is characterized in that: in step (5), then, the computing formula of trying to achieve the content of methyl eugenol in cigarette sample is as follows:

$W=\frac{(\frac{A}{\mathrm{As}}-b)\&times;\mathrm{Ms}}{a\&times;M}$

In formula: W is the content of methyl eugenol in cigarette sample, unit is that ng/ props up; A is the peak area of methyl eugenol; As is interior mark 3, the peak area of 4-(methylenedioxy) acetophenone; B is the intercept of standard working curve; Ms is interior target quality in sample solution, and unit is ng; A is the slope of standard working curve; M is the quantity of cigarette, and unit is for propping up.