CN104090043B - A kind of method measuring methyl eugenol in cigarette mainstream flue gas - Google Patents

Abstract

The invention discloses a kind of method measuring methyl eugenol in cigarette mainstream flue gas, comprise the preparation of inner mark solution, standard working solution and sample solution, gas chromatography tandem mass spectrometry method is analyzed, and the step such as the calculating of measurement result.The present invention adopts 3, 4-(methylenedioxy) acetophenone is that internal standard compound carries out quantitatively, detection method operation after optimizing is comparatively easy, respond sensitive, quantitative test is accurate, effectively reduce the interference brought because sample substrate is complicated, the chromatogram adopted and Mass Spectrometry Conditions make object have good signal response, and there is good linear dependence, detect and be limited to 0.31ng/ and prop up, relative standard deviation is 4.30%, recovery of standard addition is between 94.85% ~ 112.51%, the highly sensitive of this method is described, reproducible, the recovery is high, be suitable for the quantitative test of trace object in flue gas complex matrices.

Description

A kind of method 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 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 MethylEugenol (EugenylMethylEther) by name.No. CAS is 93-15-2, and molecular formula is C ₁₁h ₁₄o ₂, molecular weight is 178.23, colourless to micro-yellow liquid.Boiling point 249 DEG C, relative density 1.032-1.036, refractive index 1.532-1.536, flash-point 99 DEG C, is dissolved in ethanol and oils.Have the fresh and sweet pungent fragrance of cloves-fennel, like carnation breath, fragrance is more thoroughly sent out and lastingly, has the pungent fragrance of the 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, and methyl eugenol has been listed in the forbidding name list of China's " cosmetics health specification ".

For the analysis detection of methyl eugenol, abroad, alkylene benzene class aromatic substance identified and quantitative analysis in some spice berries.When analyzing the alkylene benzene class aromatic substance in plant, the instrument analytical method adopted mainly contains GC or GC-MS, and HPLC.Such as 1985, the people such as CarmanAS used GC method to carry out fast quantitative analysis to the methyl eugenol in fresh and frozen carrot, myristicin; 1988, ArcherAW used HPLC method to carry out quantitative analysis to the methyl eugenol in nutmeg and nutmeg dry hide, myristicin; When analyzing the alkylene benzene class aromatic substance in plant, the Sample Pretreatment Technique adopted, according to the difference of sample substrate, mainly contain solvent extraction (Schmelta, 1967), Solid-Phase Extraction (YatesandEngland, nineteen eighty-two), steam distillation (Lavoieetal, 1985), supercritical fluid extraction (Heikes, 1994), solid-phase microextraction (ClarkandBunch, 1997).In addition, also have the report analyzed the alkylene benzene class aromatic substance in tobacco, such as 1999, StanfillandAshley uses 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 the complicacy of flue gas matrix and the lower reason of object content, the quantitative test of alkylene benzene class aromatic substance is considered to the problem being worth challenge always.1964, Rodgman and Cook collected 20560 granule phase substances without filter tip cigarette flue gas, adopts suitable pre-treating method, was 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 Lavoieetal used GC-MS method to carry out analyzing measuring to the eugenol in cloves cigarette.Until 2000, the people such as Stephen adopt Solid-Phase Extraction-GC-MS method, 8 kinds of alkylene benzene class aromatic substances such as eugenol, methyl eugenol are comprised to the cigarette that American market is sold and carries out analysis mensuration, and analyze the impact of different filter tip degree of ventilation on alkylene benzene class aromatic substance burst size.Wherein when sample pre-treatments, according to the different content of object, be divided into two parts, i.e. extract only solid-phase extraction column (Pre-SPE) and mistake solid-phase extraction column (Post-SPE).

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 methyl eugenol contained by Herba Asari different growing stages and safrole vapor-phase chromatography.1999, the people such as Zhou Changzheng adopted internal standard method to carry out gas chromatographic analysis to the content of methyl eugenol and safrole in the Domestic Drugs root of Chinese wild ginger 12 kinds totally 24 sample volatile oil.2009, the people such as Wu Ting established with the gas chromatography that Isosorbide-5-Nitrae-dibromobenzene and phenethyl acetate are interior mapping fixed natural nuisance absinthol, esdragol and methyl eugenol content in different flavoured food product, and retention time is qualitative, inner mark method ration.2012, the people such as Chen Jianwei adopted supercritical fluid extraction and steam distillation to carry out extraction and isolation to Herba Asari root and rhizome respectively, obtained 5.5%SFE extract and 2.8% volatile oil respectively.Application GC-MS analyzes, and from Herba Asari SFE extract, identify 7 kinds of chemical compositions, account for 72.70% of extract total amount, 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 than the way of distillation 2.8 times.2012, the people such as Zhu Xiaolan establish ultrasonic solvent extracts and dispersive liquid-liquid microextraction (DLLME) combines extraction, purification pretreatment technology, 8 kinds of alkyl benzene fragrance nuisances such as absinthol, esdragol, pulegone, methyl eugenol, cumarin, 6-Methylcoumarin and ayapanin in employing GC-MS Analysis cigarette additive.The beautiful analytical approach waiting people to establish methyl eugenol in Gas Chromatography-Mass Spectrometry perfume of phase.

In sum, the methods such as GC-MS, HPLC are mainly adopted 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 Simultaneous distillation-extraction.

More than analyze the sample mainly spice berry detecting and relate to, lack the analysis and research to methyl eugenol in cigarette smoke, only report is that Stephen and Ashley in 2000 adopts Solid-Phase Extraction-GC/MS method, carries out analysis measure methyl eugenol in the cigarette smoke that American market is sold at interior 8 kinds of alkylene benzene class aromatic substances.But based on the repeated authentication to the method, on the solid-phase extraction column finding the CN bonding that methyl eugenol adopts at document, retention is strong, does not reach the object of selective elution, at all cannot accurate quantitative analysis object.

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 measuring methyl eugenol in cigarette mainstream flue gas, quick to energy, accurately to detect methyl eugenol in cigarette mainstream flue gas content, 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:

Measure a method for methyl eugenol in cigarette mainstream flue gas, comprise the following steps:

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

(2) preparation of standard working solution: take methyl eugenol as standard items, with ethanol as solvent, be prepared into Standard Stock solutions through stepwise dilution, then add a certain amount of inner mark solution respectively, be prepared into standard working solution;

(3) preparation of sample solution: gather the object in cigarette smoke, accurately adds 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: with gas chromatography tandem mass spectrometry instrument (GC-MS/MS), detection is carried out to standard working solution and sample solution and analyze; 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 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 lists in table 1;

The MRM parameter of table 1 object

(5) drafting of standard working curve and the calculating of sample result: with the ratio of methyl eugenol in standard working solution and interior target concentration for horizontal ordinate, with the ratio of methyl eugenol in chromatogram and interior target peak area for ordinate, carry out linear regression analysis, obtain standard working curve.By methyl eugenol and interior target chromatographic peak area ratio in the sample solution that records under the same terms, substitute into standard working curve, convert and try 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, obtains the interior mark storing solution that concentration is 1000 μ g/mL; 2) inner mark solution: accurately pipette mark storing solution in 1mL, with ethanol constant volume to 100mL, obtain the interior mark storing solution that concentration is 10 μ g/mL.

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

In step (3), the preparation of described sample solution specifically comprises the following steps: 1) cigarette smoking: the TPM collecting 20 cigarette according to the requirement of GB/T19609 standard; 2) sample extraction: (filter disc is answered full wafer to put into conical flask and paved the trapping that upper step obtains to be had the glass fiber filter of TPM to be positioned in 100mL conical flask, can not shred or tear up), accurately add 0.4mL inner mark solution to exist, and add 40mL normal hexane, to be placed in Clothoid type oscillator oscillation extraction 40min under 160 turns/min condition, to leave standstill a moment; 3) extract purification: accurately pipette 10mL sample extraction liquid to solid-phase extraction column (filler is the silica gel of 500mg), liquid to be extracted toward be displaced downwardly to extraction pillar top after, point adding 10mL normal hexane for 2 times carries out drip washing, drip washing speed remains on 1 ~ 2mL/min, collects all leacheates; 4) leacheate concentrates: be connected on Rotary Evaporators by the concentrated bottle filling all leacheates; at 80 DEG C, rotary evaporation under high pure nitrogen protection, or at 55 DEG C, rotary evaporation under vacuum condition, be concentrated to about 1mL; 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 the peak area of interior mark 3,4-(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 measures the content of methyl eugenol in cigarette mainstream flue gas first based on gas chromatography tandem mass spectrometry instrument, solve methyl eugenol burst size level in flue gas low, and the impact of the factor such as the Analysis interference of flue gas complex matrices on object be serious.

(2) the inventive method utilizes inner mark method ration, can accurate constant volume, and can reduce the error brought by pre-treating method reappearance and instrument precision problem.

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

Accompanying drawing explanation

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

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

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

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

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

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, obtains the interior mark storing solution that concentration is 1069 μ g/mL; Inner mark solution: accurately pipette mark storing solution in 1mL, with ethanol constant volume to 100mL, obtain the interior mark storing solution that concentration is 10.69 μ g/mL.

(2) preparation of standard working solution

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

(3) preparation of sample solution

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

(4) gas chromatography tandem mass spectrometry analysis

Standard working solution and the sample solution to be measured of getting 5 variable concentrations respectively carry out gas chromatography tandem mass spectrometry analysis.Its 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 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 lists in table 1.The extraction Ion-pair chromalography figure of safrole in standard working solution, as shown in Figure 2, wherein, upper figure is 178-> 107 ion pair, and figure below is 178-> 147 ion pair; The extraction Ion-pair chromalography figure of safrole in sample solution, as shown in Figure 3, wherein, upper figure is 178-> 107 ion pair, and figure below is 178-> 147 ion pair; The extraction Ion-pair chromalography figure of internal standard compound in standard working solution, as shown in Figure 4, wherein, upper figure is 164-> 121 ion pair, and figure below is 164-> 149 ion pair; The extraction Ion-pair chromalography figure of internal standard compound in sample solution, as shown in Figure 5, wherein, upper figure is 164-> 121 ion pair, and figure below is 164-> 149 ion pair.

The MRM parameter of table 1 object

(5) Specification Curve of Increasing and result calculate

First, with the ratio of methyl eugenol in standard working solution and interior target concentration for horizontal ordinate, with the ratio of methyl eugenol in chromatogram and interior target peak area for ordinate, carry out linear regression analysis, obtain standard working curve, get the standard working solution of least concentration, do 9 Parallel testing analyses, calculate its standard deviation, with concentration corresponding to the standard deviation of 3 times for 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 and interior target chromatographic peak area ratio in the sample solution that records under the same terms, substitute into standard working curve, try to achieve the content of methyl eugenol in cigarette sample, 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 that nanogram often props up (ng/ props up);

The peak area of A---methyl eugenol;

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

The intercept of b---standard working curve;

Ms---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.

Measure the methyl eugenol content in 5 different brands cigarette samples in the present embodiment, 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

With a cigarette sample for analytic target, adopt the method for embodiment 1 to measure the content of object, carry out Precision Experiment, measure 6 times altogether, calculate the relative standard deviation (RSD) of 6 parallel determinations respectively, measurement result sees the following form 3.Result display in table, the RSD<5% of the repeatability of this experimental technique.For trace species analysis detects, 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

With a cigarette sample for object, carry out the mark-on process of basic, normal, high three variable concentrations levels, then process according to above-mentioned standard pre-treatment step, carry out GC-MS/MS analysis again, measure the content of object, calculate its recovery of standard addition, the results are shown in following table 4.As can be seen from following table, the recovery of object is between 94.85% ~ 112.51%.On the whole, for the object that content is in several nanogram level, the method has the recovery relatively preferably.

The recovery of table 4 method

Standard solution used by the present embodiment is only described for one of them concentration, the typical curve that standard solution prepared by other concentration value obtains through gas chromatography tandem mass spectrometry analysis and regression equation same as the previously described embodiments, do not enumerating at this.Illustrated embodiment just method for a better understanding of the present invention, do not have any restriction, namely the method for said method or equivalent above-mentioned situation is all included in the protection domain of technical scheme of the present invention.

Claims (6)

1. measure a method for methyl eugenol in cigarette mainstream flue gas, it is characterized in that, comprise the steps:

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

(2) preparation of standard working solution: take methyl eugenol as standard items, with ethanol as solvent, be prepared into Standard Stock solutions through stepwise dilution, then add quantitative inner mark solution respectively, be prepared into standard working solution;

(3) preparation of sample solution: gather the object in cigarette smoke, accurately adds 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: with gas chromatography tandem mass spectrometry instrument, detection is carried out to standard working solution and sample solution and analyze; 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 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 lists in table 1;

The MRM parameter of table 1 object

(5) drafting of standard working curve and the calculating of sample result: with the ratio of methyl eugenol in standard working solution and interior target concentration for horizontal ordinate, with the ratio of methyl eugenol in chromatogram and interior target peak area for ordinate, carry out linear regression analysis, obtain standard working curve, by methyl eugenol and interior target chromatographic peak area ratio in the sample solution that records under the same terms, substitute into standard working curve, convert and try 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, obtain the interior mark storing solution that concentration is 1000 μ g/mL; 2) inner mark solution: accurately pipette mark storing solution in 1mL, with ethanol constant volume to 100mL, obtain the interior mark storing solution that concentration is 10 μ g/mL.

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 working solution specifically comprises the following steps: 1) primary standard storing solution: accurately take 100mg methyl eugenol, with ethanol constant volume to 100mL, obtain the primary standard storing solution that concentration is 1000 μ g/mL; 2) secondary standard storing solution: accurately pipette 0.5mL primary standard storing solution, with ethanol constant volume to 100mL, obtains the secondary standard storing solution that concentration is 5 μ g/mL; 3) standard working solution: accurately pipette secondary standard storing solution 10 μ L, 20 μ L, 50 μ L, 100 μ L and 200 μ L respectively, and each 100 μ L of inner mark solution, be settled in the volumetric flask of 10mL with ethanol as solvent, obtain the series standard working solution that concentration is respectively 5ng/mL, 10ng/mL, 25ng/mL, 50ng/mL and 100ng/mL, wherein, 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 collecting 20 cigarette according to the requirement of GB/T19609 standard; 2) sample extraction: the trapping that upper step obtains had the glass fiber filter of TPM to be positioned in 100mL conical flask, accurately add 0.4mL inner mark solution, and add 40mL normal hexane, to be placed in Clothoid type oscillator oscillation extraction 40min under 160 turns/min condition, to leave standstill a moment; 3) extract purification: accurately pipette 10mL sample extraction liquid to solid-phase extraction column, liquid to be extracted is behind the top being displaced downwardly to extraction pillar, and divide and add 10mL normal hexane for 2 times and carry out drip washing, drip washing speed remains on 1 ~ 2mL/min, collects all leacheates; 4) leacheate concentrates: be connected on Rotary Evaporators by the concentrated bottle filling all leacheates; at 80 DEG C, rotary evaporation under high pure nitrogen protection, or at 55 DEG C, rotary evaporation under vacuum condition, be concentrated to about 1mL; 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, it 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:

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

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 the peak area of interior mark 3,4-(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.