1. A high-efficiency detection method for volatile components in cigarette smoke is characterized by comprising the following steps:

s1, extracting smoke components, sucking and trapping the smoke of the cigarette, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments, putting the fragments into a headspace bottle, adding an internal standard solution to obtain the sample, and then putting the sample into a refrigerator for storage for later use;

s3, performing headspace solid phase microextraction, namely taking out the sample prepared in the step S2, placing the sample into a thermostat, standing and unfreezing, and then extracting by adopting full-automatic headspace solid phase microextraction HS-SPME;

and S4, performing GC-MS detection, and enabling the sample to be detected extracted in the step S3 to enter a GC-MS detection system for detection and analysis.

2. The efficient detection method for the volatile components in the cigarette smoke according to claim 1, wherein the mass ratio of the fragments to the internal standard liquid in the step S2 is 40: (1.3-2.0).

3. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 1, wherein the internal standard solution in the step S2 is [2H8] -acetophenone with a concentration of 10 μ g/mL.

4. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 1, wherein in the step S2, the Cambridge filter obtained in the step S1 is cut into small pieces with the length and the width of 1-2 cm.

5. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 1, wherein the environmental temperature for sample pretreatment in the step S2 is 0-20 ℃.

6. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 1, wherein the sample is stored in a refrigerator at-20 ℃ for later use in step S2.

7. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 1, wherein the heating temperature in the step S3 is 50-80 ℃.

8. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 1, wherein the sample prepared in the step S2 is taken out from the step S3 and is placed in a constant temperature box for standing at 16-20 ℃.

9. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 8, wherein the sample prepared in the step S2 is taken out and placed in an incubator for standing for 25-35min in the step S3.

10. The method for efficiently detecting the volatile components in the cigarette smoke according to claim 1, wherein the step S4 further comprises the following steps: a normal paraffin mixture solution with a concentration of 10ppm was prepared and subjected to GC-MS analysis as an external standard together with the sample extracted in step S3.

Background

Cigarette combustion is a complex chemical system, except for gas phase substances (such as carbon monoxide, water, methane and other lower hydrocarbon compounds) generated by full combustion and cracking of organic matters, due to insufficient oxygen supply in partial areas, a plurality of complex chemical changes occur in an anoxic state, volatile substances (such as semi-volatile 5-membered ring and 6-membered ring nitrogen-heterocyclic compounds) in tobacco shreds are volatilized into smoke gas flow, and terpenes, saccharides, amino acids, celluloses and other components in tobacco are subjected to thermal decomposition, thermal synthesis, dry distillation, polymerization, condensation, free radical and other reactions to generate volatile and semi-volatile gases. The smoke components generated after the cigarette is burnt are very complex, and comprise carboxylic acids, lipids, aldehydes, ketones, nicotine, alkane and the like, and the specific chemical substances are more than 4000. Wherein the harmful substances affect the sleep quality and fertility of smokers, and cause chronic bronchitis. Osteoporosis and other health problems, and even serious diseases such as coronary heart disease, cancer and the like. Along with the improvement of health consciousness of people, the attention to harmful substances in smoke generated by cigarette combustion is increased, and a plurality of products capable of reducing the harmfulness of cigarettes are designed. For example, the external filter cigarette holder directly blocks substances such as tar in smoke through micropores, but the substances block harmful substances only through an absorption means, and the harm of the cigarette to human bodies cannot be reduced substantially.

Therefore, before the cigarette leaves factory, the analysis and determination of the composition of the components in the smoke has very important significance for improving the quality of the tobacco leaves and promoting the sustainable development of the tobacco leaves. In the prior art, the detection method of cigarette smoke components is mainly a GC-MS method, but most of the methods have the problems of complex operation, small detection range and the like. Therefore, a method for measuring chemical components in smoke with high speed, high efficiency, simple operation and wide detection range is urgently needed.

Chinese patent (CN 106556665A) discloses a method for simultaneously measuring 14 sour ingredients in main stream smoke of cigarettes, which comprises the following steps: 1) after particulate matters of mainstream smoke of the cigarettes are captured by using a Cambridge filter, the Cambridge filter is transferred into an extraction container, an acetone solution is added, an internal standard solution is added for oscillation extraction, and a supernatant is taken, added with a derivatization reagent and heated to obtain a smoke sample; 2) and (3) detecting the flue gas sample by adopting GC-MS (gas chromatography-Mass spectrometer), and detecting the content of 14 sour components by an internal standard quantitative method. The method adopts solvent extraction-gas chromatography-mass spectrometry combined method, can simultaneously determine the content of 14 sour components in cigarette mainstream smoke, and is suitable for determining the content of the sour components in cigarette smoke. However, the method is complicated to operate, an organic solvent is needed as a solvent for back extraction, and in the process, the method of shaking, shaking or heating is adopted, so that volatile substances are easily lost.

Chinese patent (CN 108680694A) discloses a method for measuring volatile chemical components in smoke by using an infrared mirror reflection furnace-headspace gas chromatography-mass spectrometry, which comprises the following steps: (A) placing the tobacco sample and the glass fiber filter disc under constant temperature and humidity for balancing for 48 hours; (B) weighing not less than 0.50g of the tobacco leaf sample obtained in the step A, placing the tobacco leaf sample into a quartz tube, heating by using infrared rays, carrying out temperature rise program heating, introducing air into the quartz tube during heating to simulate the combustion process of the tobacco leaves, stopping infrared ray heating after a period of time, introducing nitrogen into the quartz tube to simulate the smoldering process of the tobacco leaves, trapping chemical components in smoke by using a glass fiber filter disc, and repeatedly carrying out the steps of simulating combustion and simulating smoldering for a plurality of times; (C) and (3) putting the glass fiber filter disc into a headspace bottle, and performing headspace gas chromatography-mass spectrometry combined analysis. The method can make the tobacco shred consumption reach or even exceed 1 cigarette tobacco leaf raw material, and ensure accurate analysis of trace amount of volatile chemical components in tobacco leaf smoke. However, the method adopts an infrared mirror reflection furnace to simulate the combustion process of the flue gas, and only 100 kinds of volatile compounds can be obtained by measurement according to the method, so that the requirement of wide detection of flue gas components cannot be met.

Disclosure of Invention

In order to solve the problems, the invention provides a high-efficiency detection method for volatile components in cigarette smoke, which specifically comprises the steps of smoke component extraction, sample pretreatment, headspace solid phase microextraction, GC-MS detection and the like.

The technical scheme provided by the invention is as follows: a high-efficiency detection method for volatile components in cigarette smoke comprises the following steps:

s1, extracting smoke components, sucking and trapping the smoke of the cigarette, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments, putting the fragments into a headspace bottle, adding an internal standard solution to obtain the sample, and then putting the sample into a refrigerator for storage for later use;

s3, performing headspace solid phase microextraction, namely taking out the sample prepared in the step S2, placing the sample into a thermostat, standing and unfreezing, and then extracting by adopting full-automatic headspace solid phase microextraction HS-SPME;

and S4, performing GC-MS detection, and enabling the sample to be detected extracted in the step S3 to enter a GC-MS detection system for detection and analysis.

Further, the mass ratio of the fragments to the internal standard solution in the step S2 is 40: (1.3-2.0).

Further, the internal standard solution in the step S2 is [2H8] -acetophenone with the concentration of 10 μ g/mL.

Further, in the step S2, the cambridge filter obtained in the step S1 is cut into small pieces each having a length and a width of 1-2 cm.

Further, the environmental temperature of the sample pretreatment in the step S2 is 0-20 ℃.

Further, in the step S2, the sample is stored in a refrigerator at-20 ℃ for further use.

Further, the heating temperature during the extraction in the step S3 is 50-80 ℃.

Further, in the step S3, the sample prepared in the step S2 is taken out and placed in an incubator for standing at 16-20 ℃.

Further, in the step S3, the sample prepared in the step S2 is taken out and placed in an incubator for standing for 25-35 min.

Further, the step S4 includes the following steps: a normal paraffin mixture solution with a concentration of 10ppm was prepared and subjected to GC-MS analysis as an external standard together with the sample extracted in step S3.

The internal standard solution is a method for accurate quantification in chromatographic analysis, a certain weight of pure substances are added into a certain amount of sample mixture to be analyzed as internal standard substances, and the content of the component to be detected is calculated according to the mass ratio of the sample to be detected to the internal standard substances, the ratio of corresponding chromatographic peak areas and relative correction factors.

The general internal standard solution selection criteria are as follows:

a. the original sample contains no components

b. The retention time of the sample should be close to but not overlapped with that of the sample

c. Is a high purity standard substance, or a substance with a known content

d. Has certain chemical stability under given chromatographic conditions

Therefore, the invention selects the [2H8] -acetophenone as the isotope internal standard substance, and the accuracy is very high.

The invention has the beneficial effects that:

1. the invention provides a high-efficiency detection method of volatile components in cigarette smoke, which specifically comprises the steps of smoke component extraction, sample pretreatment, headspace solid phase microextraction, GC-MS detection and the like, wherein more than 500 volatile components can be obtained by detecting smoke through the method, and the method has the advantages of wide detection range, simple and convenient operation, rapidness and high efficiency. The detection method provided by the invention can detect smoke components more comprehensively, quickly and efficiently, and can judge the quality of cigarettes better.

2. According to the efficient detection method for volatile components in cigarette smoke, provided by the invention, the Cambridge filter fragment after smoke collection is directly subjected to headspace solid phase microextraction, other intermediate treatment steps are not required, smoke components are retained to the maximum extent, the loss of volatile components in the treatment process can be reduced, the detection of more than 500 volatile components in the smoke can be realized, and the detection range of disposable smoke is widest based on the GC-MS technology at present.

Drawings

FIG. 1 is a flow chart of a method for efficiently detecting volatile components in cigarette smoke according to an embodiment of the present invention.

Detailed Description

In order to more clearly illustrate the embodiments of the present invention and the technical solutions in the prior art, the following will describe the specific embodiments of the present invention with reference to the accompanying drawings.

It is obvious that the drawings in the following description are only some examples of the invention, and it is obvious to a person skilled in the art that other drawings and other embodiments can be obtained from these drawings without inventive effort, and the invention is not limited to this example.

In the embodiment of the invention, SPME sample injection parameters are as follows: aging temperature: 250 ℃; aging time: 5 min; heating temperature: 60 ℃; heating time: 10 min; adsorption time: 20 min; analysis time: 5 min; aging time after sample introduction: and 5 min.

The chromatographic collection conditions were: shunting mode: no flow diversion; carrier gas: he; a chromatographic column: DB-5MS (30m x 0.25mm x 0.25 μm); column flow rate: 1.0 mL/min; column box temperature program: the temperature was maintained at 40 ℃ for 5 minutes, and then the temperature was raised to 280 ℃ (6 ℃/min) for 5 minutes. The mass spectrum acquisition conditions are as follows: temperature of a front sample inlet: 250 ℃; transmission line temperature: 280 ℃; ion source temperature: 230 ℃; quadrupole temperature: 150 ℃; ionization voltage: 70 eV; solvent retardation: and 5 min. See table 1 for details.

Table 1: collecting conditions of chromatographic mass spectrum

Item	Parameter(s)
		Sample volume	-
Shunting Mode (Front Inlet Mode)	splitless
		Carrier Gas (Carrier Gas)	Helium
Chromatographic Column (Column)	DB-5MS (30m x 0.25mm x 0.25μm)
		Column Flow rate (Column Flow)	1.0mL/min
Column box Temperature program (Oven Temperature Ramp)	40°C hold on 5 min, raised to 280°C at a rate of 6 °C/min, hold for 5 min
		Front Injection Temperature (Front Injection Temperature)	250°C
Transmission Line Temperature (Transfer Line Temperature)	280°C
		Ion Source Temperature (Ion Source Temperature)	230°C
Four-lever Temperature (Quad Temperature)	150°C
		Ionization voltage (Electron Energy)	70 eV
Solvent Delay (Solvent Delay)	5 min

In the embodiment of the invention, the specific steps of smoking and trapping of smoke are carried out according to the method specified in GB/T19609-2004, so that the mainstream smoke of the cigarette to be detected completely passes through the Cambridge filter disc, the Cambridge filter disc is used for trapping volatile components in the smoke, then the Cambridge filter disc collecting the smoke is detected, and classification numbering is carried out according to the difference of the smoke.

In the present example, 12 samples were prepared in total and the metabolic studies were performed in 4 groups, wherein each group had 3 biological replicates. The grouping of each sample and the corresponding information are shown in table 2. The 12 samples were subjected to the component detection using the method provided by the present invention, and the 12 samples corresponded to examples 1 to 12, respectively.

Table 2: sample number information

Examples	Sample grouping	Sample name	Sample description
				Example 1	CZGZ	CZGZ-1	Flue gas
Example 2	CZGZ	CZGZ-2	Flue gas
				Example 3	CZGZ	CZGZ-3	Flue gas
Example 4	CZG4	CZG4-1	Flue gas
				Example 5	CZG4	CZG4-2	Flue gas
Example 6	CZG4	CZG4-3	Flue gas
				Example 7	CZGZ7	CZGZ7-1	Flue gas
Example 8	CZGZ7	CZGZ7-2	Flue gas
				Example 9	CZGZ7	CZGZ7-3	Flue gas
Example 10	CZGZ9	CZGZ9-1	Flue gas
				Example 11	CZGZ9	CZGZ9-2	Flue gas
Example 12	CZGZ9	CZGZ9-3	Flue gas

The specific embodiment of the invention is as follows:

example 1

S1, extracting smoke components, namely collecting the smoke CZGZ-1, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 20 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 25min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 50 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 2

S1, extracting smoke components, namely collecting the smoke CZGZ-2, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1.5cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 25 mu L of internal standard liquid to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard liquid is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 30min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 65 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 3

S1, extracting smoke components, namely collecting the smoke CZGZ-3, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 2cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 30 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 35min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 80 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 4

S1, extracting smoke components, namely collecting the smoke CZG4-1, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 20 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 25min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 50 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 5

S1, extracting smoke components, namely collecting the smoke CZG4-2, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1.5cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 25 mu L of internal standard liquid to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard liquid is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 30min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 65 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 6

S1, extracting smoke components, namely collecting the smoke CZG4-3, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 2cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 30 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 35min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 80 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 7

S1, extracting smoke components, namely collecting the smoke CZGZ7-1, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 20 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 25min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 50 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 8

S1, extracting smoke components, namely collecting the smoke CZGZ7-2, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1.5cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 25 mu L of internal standard liquid to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard liquid is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 30min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 65 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 9

S1, extracting smoke components, namely collecting the smoke CZGZ7-3, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 2cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 30 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 35min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 80 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 10

S1, extracting smoke components, namely collecting the smoke CZGZ9-1, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 20 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 25min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 50 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 11

S1, extracting smoke components, namely collecting the smoke CZGZ9-2, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 1.5cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 25 mu L of internal standard liquid to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard liquid is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 30min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 65 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Example 12

S1, extracting smoke components, namely collecting the smoke CZGZ9-3, and enabling the mainstream smoke of the cigarette to be detected to pass through a Cambridge filter disc;

s2, preprocessing a sample, cutting the Cambridge filter disc obtained in the step S1 into small fragments with the length and width of 2cm on an ice block, taking 0.4g of the fragments into a headspace bottle, quickly adding 30 mu L of internal standard solution to obtain a sample to be detected, screwing a bottle cap, and then placing the sample to be detected into a refrigerator for storage at-20 ℃ for later use, wherein the internal standard solution is [2H8] -acetophenone with the concentration of 10 mu g/mL;

s3, performing headspace solid-phase microextraction, namely taking out the sample to be detected prepared in the step S2, placing the sample into a thermostat, standing for 35min for thawing, and then extracting by adopting full-automatic headspace solid-phase microextraction HS-SPME, wherein the heating temperature is 80 ℃ during extraction;

and S4, performing GC-MS detection, and performing detection analysis on the sample extracted in the step S3 in a GC-MS detection system. And preparing normal alkane mixed liquor with the concentration of 10ppm as an external standard and carrying out GC-MS analysis together with a sample.

Data processing method

The original data files obtained from the GC-MS analysis in examples 1-12 were first subjected to peak extraction by MassHunter software (Agilent) to obtain information on mass-to-charge ratio, retention time, and peak area of characteristic peaks, and then the data were subjected to statistical analysis. To better analyze the data, embodiments of the present invention perform a series of preparations and arrangements on the raw data. The method mainly comprises the following steps:

(1) calculating a retention index;

(2) filtering the single Peak, and only retaining Peak area data with a single null value of no more than 50% or with null values of no more than 50% in all groups;

(3) data normalization (normalization) using an internal standard.

Method for analyzing results

Volatile Organic Compounds (VOCs) are generally understood to be various organic compounds having a boiling point of from 50 ℃ to 260 ℃ at atmospheric pressure. VOCs can be further classified into alkanes, aromatic hydrocarbons, esters, aldehydes, and others according to their chemical structures. Benzene, toluene, xylene, styrene, phenethyl alcohol, chloroform, propyl butyrate, 2-nonanone, 2-octanone, etc. are common. Plants release a large amount of volatile organic compounds, mainly isoprenoids and monoterpenes, to the atmosphere in physiological processes. Plant-derived volatile organic compounds (BVOCs) have an extremely important ecological function. BVOCs are bio-signal substances that can transmit signals or information between plant tissues-organs, plant-plants and plant-animals to warn other organisms in the vicinity, resist environmental stress, prevent plant diseases and insect pests, etc. Moreover BVOCs affect the carbon cycle of the terrestrial ecosystem and the atmospheric photochemical processes. The MEIWEI volatile metabolism group adopts NIST database to detect terpenes, benzenoids, phenylpropanoids, alcohols, fatty acid derivatives, alkanes, ketones, esters, aldehydes, etc.

Therefore, the experimental data of the samples to be tested obtained in examples 1 to 12 are collated and analyzed based on the NIST database and in combination with the data processing method.

And (4) reporting the identified compound with the matching degree of more than 80% by NIST spectral library retrieval, normal paraffin retention time analysis and artificial analysis of experimental data. Opening a machine profile file under a sample to be detected by MassHunter quantitative software, integrating and correcting chromatographic peaks, wherein the peak Area (Area) of each chromatographic peak represents the relative content of a corresponding substance, and finally exporting integral data of all chromatographic peak areas for storage. 533 volatile substances are obtained by the co-detection in the embodiment of the invention, wherein the first five substances with the largest variety are 168 heterocyclic compounds, 86 aromatic hydrocarbons, 50 esters, 46 ketones, 32 alkanes, and other olefins, terpenes, amines, alcohols, phenols, sulfides, etc. Table 3 lists the major metabolites detected:

table 3: statistical table of metabolite quantities

Substance(s)	CAS	Molecular formula	Precise molecular mass
				1, 3-dioxolan-2-ones	96-49-1	C3H4O3	88.016
(E) -3-penten-2-one	3102-33-8	C5H8O	84.058
				(dimethylamino) acetonitrile	926-64-7	C4H8N2	84.069
Pyridine compound	110-86-1	C5H5N	79.042
				1-hydroxy-2-butanone	5077-67-8	C4H8O2	88.052
Dihydro-2-methyl-3 (2H) -furanones	3188-00-9	C5H8O2	100.052
				2-methyl radicalPyridine compound	109-06-8	C6H7N	93.058
Methylpyrazine	109-08-0	C5H6N2	94.053
				1, 4-dimethylpyrazole	1072-68-0	C5H8N2	96.069
3-Furanylmethanol	4412-91-3	C5H6O2	98.037
				3-methylpyridine	108-99-6	C6H7N	93.058
(Aminoiminomethyl) -urea	141-83-3	C2H6N4O	102.054
				4-cyclopentene-1, 3-dione	930-60-9	C5H4O2	96.021
2, 6-lutidine	108-48-5	C7H9N	107.073
				2-methyl butyric acid	116-53-0	C5H10O2	102.068
3-methyl-2-cyclopenten-1-one	2758-18-1	C6H8O	96.058
				2-ethylpyridines	100-71-0	C7H9N	107.073
2, 6-dimethylpyrazine	108-50-9	C6H8N2	108.069
				2-butenoic acid, 3-methyl-2-methylene-3-butenyl ester	76003-38-8	C10H14O2	166.099
2, 5-hexanedione	110-13-4	C6H10O2	114.068
				2, 5-dimethylpyridine	589-93-5	C7H9N	107.073
2, 3-dimethyl-2-cyclopenten-1-one	1121-05-7	C7H10O	110.073
				3, 4-dimethyl-2-cyclopenten-1-one	30434-64-1	C7H10O	110.073
2, 3-dimethylpyridine	583-61-9	C7H9N	107.073
				N, N-diethyl-3-butan-2-amine	37969-64-5	C8H17N	127.136
Propyl benzene	103-65-1	C9H12	120.094
				Cyanoacetic acid methyl ester	105-34-0	C4H5NO2	99.032
2, 2-dimethyl-4-ethynyl-3, 6-dihydro-2H-pyran	42491-40-7	C9H12O	136.089
				1H-imidazole-4-carbaldehyde	3034-50-2	C4H4N2O	96.032
4-vinylpyridines	100-43-6	C7H7N	105.058
				4-hexen-3-ones	2497-21-4	C6H10O	98.073
4H-pyran-4-ones	108-97-4	C5H4O2	96.021
				Alpha, alpha-dimethyl benzyl alcohol	617-94-7	C9H12O	136.089
Benzonitrile	100-47-0	C7H5N	103.042
				6-methyl-5-hept-2-one	110-93-0	C8H14O	126.104
1-methylimidazole-5-carbaldehyde	39021-62-0	C5H6N2O	110.048
				2,2', 5,5' -tetrahydro-2, 2' -bifuran	98869-92-2	C8H10O2	138.068
2-Furanylmethanol, acetate salt	623-17-6	C7H8O3	140.047
				1,2, 3-trimethylbenzene	526-73-8	C9H12	120.094
Phosphorous acid vinyl ester	58402-90-7	C2H5O3P	107.998
				Acetochlor alcohol	113-18-8	C7H9ClO	144.034
Squalene	111-02-4	C30H50	410.391
				2,3' -bipyridine	581-50-0	C10H8N2	156.069
Dihydroactinidiolide	17092-92-1	C11H16O2	180.115

The above-described aspects may be implemented individually or in various combinations, and such variations are within the scope of the present invention.

It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.

The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the specific embodiments of the invention be limited to these descriptions. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.