Method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol and application thereof

Abstract

The invention provides a method for detecting volatile and semi-volatile organic compounds in aerosol of a heated cigarette and application of the method. The method for detecting the volatile and semi-volatile organic compounds in the aerosol of the heated cigarette comprises the following steps: (1) smoking the heated cigarette to be detected, and trapping the aerosol by using a filter disc and an adsorbent; (2) mixing the filter disc capturing the aerosol, the adsorbent, an internal standard and a solvent, and extracting to obtain an extract; and (3) carrying out gas chromatography-tandem mass spectrometry detection on the extract, and obtaining the content of volatile and semi-volatile organic compounds in the heated cigarette to be detected according to the detection result. The method provided by the invention can simultaneously separate and analyze the content of volatile and semi-volatile organic compounds in various aerosols, improves the detection flux, improves the analysis efficiency, simplifies the pretreatment process, has the advantages of high flux, good peak shape, high separation degree, high sensitivity, good recovery rate and the like, and is suitable for industrial production. The method is suitable for analysis and determination of main volatile and semi-volatile organic compounds in heated cigarette aerosol.

Description

Technical Field

[0001] This invention belongs to the field of tobacco component analysis technology, specifically relating to a method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol and its application. Background Technology

[0002] Heated tobacco products are an important category of new tobacco products, primarily using an external heat source to heat tobacco. Based on different heating principles, they can be divided into electrically heated, fuel-heated, and physicochemically heated types. Unlike traditional cigarettes where tobacco is burned, heated tobacco products do not require combustion. Flavor components and nicotine are released through volatilization, providing consumers with a smoking experience similar to traditional cigarettes, effectively reducing the release of harmful components produced by the high-temperature combustion of tobacco. Volatile and semi-volatile organic compounds (VOCs) are a class of harmful components in heated cigarette smoke, listed as key concern components by many countries and international organizations. Among them, 1,3-butadiene, isoprene, acrylonitrile, benzene, and toluene are identified as carcinogens by the International Agency for Research on Cancer (IARC); pyridine, styrene, and quinoline are highly irritating; and ethylene oxide, propylene oxide, and glycidyl can damage the eyes, nose, and mucous membranes. Therefore, it is essential to establish methods for simultaneous detection and accurate quantification to assess the release of harmful substances in heated cigarettes. Currently, the main methods for detecting volatile organic compounds include gas chromatography, liquid chromatography, and gas chromatography-mass spectrometry (GC-MS). Gas chromatography has low throughput and many interfering factors; liquid chromatography is mainly used to analyze thermally unstable semi-volatile organic compounds, but it has low throughput and low sensitivity; GC-MS has improved sensitivity and accuracy compared to gas chromatography, but it is still lower than gas chromatography-tandem mass spectrometry, especially when detecting trace components in complex samples.

[0003] Therefore, how to provide a gas chromatography-tandem mass spectrometry (GC-MS / MS) method that can simultaneously detect multiple volatile and semi-volatile organic compounds, achieving high throughput, high sensitivity, good accuracy and reproducibility, and strong anti-interference ability, and providing a basic reference for assessing the health effects of harmful components in heated cigarette smoke on humans, has become an urgent problem to be solved. Summary of the Invention

[0004] To address the shortcomings of existing technologies, the present invention aims to provide a method for detecting volatile and semi-volatile organic compounds (VOCs) in heated cigarette aerosols and its applications. The method provided by this invention can simultaneously separate and analyze the content of VOCs and semi-volatile organic compounds in multiple aerosols, increasing detection throughput, improving analytical efficiency, and simplifying the pretreatment process. It has advantages such as high throughput, good peak shape, high resolution, high sensitivity, and good recovery rate, and is suitable for the analysis and determination of major VOCs and semi-volatile organic compounds in heated cigarette aerosols.

[0005] To achieve this objective, the present invention adopts the following technical solution: On one hand, the present invention provides a method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol, the method comprising the following steps: (1) The heated cigarette to be tested is inhaled, and the aerosol is captured using a filter and an adsorbent; (2) The filter containing the aerosol and the adsorbent are mixed with the internal standard and solvent for extraction to obtain the extract; (3) The extract was subjected to gas chromatography-tandem mass spectrometry detection, and the content of volatile and semi-volatile organic compounds in the heated cigarette to be tested was obtained based on the detection results and the standard curve.

[0006] The volatile and semi-volatile organic compounds include any one or a combination of at least two of the following: vinyl chloride, 1,3-butadiene, isoprene, ethylene oxide, propylene oxide, benzene, acrylonitrile, toluene, ethylbenzene, p-xylene, m-xylene, o-xylene, pyridine, styrene, glycidyl ether, acetamide, nitrobenzene, acrylamide, or quinoline.

[0007] The above method can simultaneously separate and analyze the content of volatile and semi-volatile organic compounds in multiple (19 kinds) aerosols, improve detection throughput, enhance analytical efficiency, and simplify the pretreatment process. It has the advantages of high throughput, good peak shape, high separation, high sensitivity, and good recovery rate, and is suitable for the analysis and determination of major volatile and semi-volatile organic compounds in heated cigarette aerosols.

[0008] Preferably, the volatile or semi-volatile organic compounds include vinyl chloride, 1,3-butadiene, isoprene, ethylene oxide, propylene oxide, benzene, acrylonitrile, toluene, ethylbenzene, p-xylene, m-xylene, o-xylene, pyridine, styrene, glycidyl ether, acetamide, nitrobenzene, acrylamide, and quinoline.

[0009] Preferably, the internal standard in step (2) includes benzene-d6.

[0010] Preferably, the solvent in step (2) is a carbon disulfide-methanol mixed solvent, wherein the volume ratio of carbon disulfide to methanol is 1:(3-5), such as 1:3, 1:3.5, 1:4, 1:4.5 or 1:5, but not limited to the values ​​listed above. Other unlisted values ​​within the above range are also applicable.

[0011] Preferably, the chromatographic column in the gas chromatography-tandem mass spectrometry step (3) is an Agilent DB-WAX UI column.

[0012] Preferably, in step (3), the temperature program for gas chromatography-tandem mass spectrometry is as follows: The initial temperature is 35-43℃, held for 4-6 minutes, then increased to 95-105℃ at 3-5℃ / min, then increased to 220-240℃ at 45-55℃ / min, held for 5-7 minutes, and then run for 2-4 minutes.

[0013] Preferably, in the gas chromatography-tandem mass spectrometry described in step (3), the carrier gas flow rate is 0.8-1.2 mL / min, such as 0.8 mL / min, 0.9 mL / min, 1 mL / min, 1.1 mL / min or 1.2 mL / min, but not limited to the values ​​listed above. Other unlisted values ​​within the above range are also applicable.

[0014] The specific reagents and parameter controls described above can effectively separate various volatile and semi-volatile organic compounds, thereby improving detection results.

[0015] Preferably, the standard curve in step (3) is prepared by a method comprising the following steps: A series of standard working solutions containing standard volatile and semi-volatile organic compounds were prepared and mixed with internal standards for gas chromatography-tandem mass spectrometry detection. A standard curve was constructed with the ratio of the quantitative ion peak area of ​​volatile and semi-volatile organic compounds in each standard working solution to that of the internal standard as the ordinate and the content of volatile and semi-volatile organic compounds in each standard working solution as the abscissa.

[0016] On the other hand, the present invention also provides the application of the method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol as described above in the analysis of harmful components in heated cigarettes.

[0017] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosols. It can simultaneously separate and analyze the content of volatile and semi-volatile organic compounds in multiple aerosols, improving detection throughput, enhancing analytical efficiency, and simplifying the pretreatment process. It has advantages such as high throughput, good peak shape, high separation, high sensitivity, and good recovery rate, and is suitable for the analysis and determination of major volatile and semi-volatile organic compounds in heated cigarette aerosols. Attached Figure Description

[0018] Figure 1 This is a graph showing the test results of the standard working solution in Example 1; Figure 2 This is a graph showing the detection results of the DB-35 MS column in Example 2; Figure 3 This is a graph showing the detection results of the DB-624 UI chromatographic column in Example 2; Figure 4This is a graph showing the detection results of the HP-INNOWAX chromatographic column in Example 2; Figure 5 This is a graph showing the detection results when the initial temperature was 35°C in Example 3; Among them, 1-1,3-butadiene; 2-chloroethylene; 3-isoprene; 4-ethylene oxide; 5-propylene oxide; 6-benzene / benzene-d6 (internal standard); 7-acrylonitrile; 8-toluene; 9-ethylbenzene; 10-p-xylene; 11-m-xylene; 12-o-xylene; 13-pyridine; 14-styrene; 15-glycidyl; 16-acetamide; 17-nitrobenzene; 18-acrylamide; 19-quinoline. Detailed Implementation

[0019] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention in any way.

[0020] Example 1: A method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosols (1) Instruments and reagents Instruments: Gas chromatography-tandem mass spectrometry (GC-MS / MS) system was Agilent 8890-7010B (USA); qualitative analysis and quantitative processing software was Agilent MassHunter Qualitative Analysis 10.0 and Quant-My-Way; electronic balance (AE163, Mettler, Switzerland, sensitivity: 0.0001 g); Talboys digital display multi-tube vortex mixer.

[0021] Reagents and consumables: Isoprene, ethylene oxide, propylene oxide, benzene, acrylonitrile, toluene, ethylbenzene, p-xylene, m-xylene, o-xylene, pyridine, styrene, glycidyl ether, acetamide, nitrobenzene, acrylamide, and quinoline were all purchased from SIGMA with a purity greater than 98.0%. Vinyl chloride and 1,3-butadiene were purchased from Tanmo Quality Inspection. Carbon disulfide was purchased from Aladdin Biochemical Technology. Methanol was purchased from TEDIA, USA (chromatographic grade).

[0022] (2) Instrument operating conditions Chromatographic conditions: Agilent DB-WAX UI column (60 m × 0.25 mm × 0.25 μm); injection port temperature: 250℃; transfer line temperature: 230℃; carrier gas: helium (≥99.999%); carrier gas flow rate: 1.0 mL / min; collision gas: nitrogen; constant flow mode; split ratio: 20:1; injection volume: 1 μL; solvent delay: 3.5 min; temperature program: initial temperature 40℃, hold for 5 min, increase to 100℃ at 4℃ / min, increase to 230℃ at 50℃ / min, hold for 6 min, total run time 28.6 min, followed by a 3 min run.

[0023] Mass spectrometry conditions: Ion source temperature (TEM): 230℃; Quadrupole temperature: 150℃; Ionization energy: 70 eV; Scan mode: full scan mode (Scan) and multiple reaction monitoring mode (MRM).

[0024] The MRM mass spectrometry detection parameters for the target compound and the internal standard are as follows: Note: "*" indicates quantitative ions.

[0025] (3) Sample pretreatment Using a smoking machine and the ISO recommended suction mode (35 mL suction volume, 2 s suction time, and 60 s suction interval), heated cigarette samples were aspirated. Cambridge filters (44 mm) and CX-572 adsorbent were used to capture the target analytes in the aerosols. The Cambridge filters and CX-572 particles containing the aerosols were placed in an extraction flask, and 100 μL of internal standard solution and 10 mL of a carbon disulfide-methanol mixture (1:4, v / v) were added. After shaking and extraction for 30 min, the mixture was filtered through a 0.22 μm organic filter membrane and transferred to a chromatographic analysis flask for analysis.

[0026] (4) Preparation of standard working solution ① Preparation of internal standard solution: Accurately weigh 50.0 mg of benzene-d6 into a 50 mL brown volumetric flask, and dilute to volume with methanol to prepare an internal standard stock solution with a concentration of 1 mg / mL. Take 1 mL of the benzene-d6 internal standard stock solution and dilute to 10 mL with methanol to obtain an internal standard solution with a concentration of 100 μg / mL.

[0027] ② Preparation of primary mixed standard stock solution Accurately weigh or transfer appropriate amounts of vinyl chloride, 1,3-butadiene, isoprene, ethylene oxide, propylene oxide, benzene, acrylonitrile, toluene, ethylbenzene, p-xylene, m-xylene, o-xylene, pyridine, styrene, glycidyl ether, acetamide, nitrobenzene, acrylamide, and quinoline into 50 mL amber volumetric flasks. Dilute to the mark with methanol to prepare a mixed standard stock solution with a concentration of 1 mg / mL. This solution should be stored at -80°C protected from light.

[0028] ③ Preparation of secondary mixed standard stock solution Accurately transfer 1 mL of the primary mixed standard stock solution into a 10 mL brown volumetric flask, and dilute to the mark with methanol to prepare a secondary mixed standard stock solution with a concentration of 100 μg / mL.

[0029] ④ Preparation of standard working solutions Accurately transfer 1 μL, 2 μL, 5 μL, 10 μL, 20 μL, 50 μL, 100 μL, 200 μL, and 500 μL of the secondary mixed standard stock solution into different 10 mL brown volumetric flasks. Then accurately add 100 μL of internal standard solution to each flask and dilute to the mark with a carbon disulfide-methanol mixed solution (volume ratio 1:4) to obtain a series of standard working solutions with 9 different concentrations.

[0030] (5) Sample determination The standard working solution obtained in step (4) and the sample solution obtained in step (3) were analyzed by GC-MS / MS, respectively. The chromatograms of the target analyte (5 μg / mL) and its internal standard in the standard working solution are shown in the figure. Figure 1 (Among them, 1-1,3-butadiene; 2-chloroethylene; 3-isoprene; 4-ethylene oxide; 5-propylene oxide; 6-benzene / benzene-d6 (internal standard); 7-acrylonitrile; 8-toluene; 9-ethylbenzene; 10-p-xylene; 11-m-xylene; 12-o-xylene; 13-pyridine; 14-styrene; 15-glycidyl; 16-acetamide; 17-nitrobenzene; 18-acrylamide; 19-quinoline, the same below). A standard curve was prepared with the ratio of the quantitative ion peak area of ​​the target substance to the internal standard in each standard working solution as the ordinate and the content of the target substance in each standard working solution as the ordinate. The analysis results of step (3) were substituted into the standard curve to obtain the content of the target substance in the test solution, and then the content of each target substance in the sample was further calculated.

[0031] (6) Method Validation The limit of detection (LOD) of this method was calculated based on the lowest-level standard working solution and a signal-to-noise ratio of 3. The recoveries were calculated at three spiking levels (low, medium, and high), with six parallel samples added at each level. The precision was calculated based on the parallel test results. The linear range, linear coefficient, LOD, Quantitation limit, average recoveries, and average precision of this method are shown in the table below. (7) Analysis of actual samples Based on the above determination method, 11 heated cigarette samples were selected, and the content of the target substance was measured as shown in the table below: Example 2: Column Selection Experiment Considering the chemical properties of the target analytes, medium-grade and highly polar chromatographic columns were mainly selected for the study, namely Agilent's DB-35 MS (30 m × 0.25 mm × 0.25 μm), DB-624 UI (30 m × 0.25 mm × 0.25 μm), HP-INNOWAX (30 m × 0.25 mm × 0.25 μm), and DB-WAX UI (60 m × 0.25 mm × 0.25 μm) (Example 1). The remaining procedures were the same as in Example 1.

[0032] Research has found that: ① On a DB-35 MS column, vinyl chloride and 1,3-butadiene do not show peaks; ethylene oxide and propylene oxide show very low responses, and the peaks of p-xylene and m-xylene overlap (e.g.) Figure 2 (As shown).

[0033] ② DB-624 UI column, no peak for 1,3-butadiene, low response and no baseline separation for vinyl chloride and ethylene oxide, overlapping peaks for pyridine and glycidyl, and overlapping peaks for p-xylene and m-xylene, and o-xylene and styrene (e.g.) Figure 3 (As shown).

[0034] ③ On the HP-INNOWAX column, the 1,3-butadiene and vinyl chloride lines were not separated, and the o-xylene and pyridine peaks overlapped. (e.g.) Figure 4 (As shown).

[0035] ④ The DB-WAX UI column can separate all 19 compounds, achieving the required resolution, with good peak shape and stability (e.g., Figure 1 (As shown).

[0036] Example 3: Experiment on the selection of heating program To investigate the effect of the heating program on the detection method, the effects of three heating programs on resolution and peak shape were examined. Three initial temperatures were set: 35°C, 40°C, and 45°C. The remaining procedures were the same as in Example 1.

[0037] Research has found that: At an initial temperature of 35℃, all 19 target analytes showed good peak elution, and the separation degree was able to reach baseline separation (see...). Figure 5 ).

[0038] With an initial temperature of 40℃, the overall peak elution time of the target analytes is shifted forward, the analysis time is shortened, and baseline separation is achieved for all target analytes.

[0039] At an initial temperature of 45°C, baseline separation of pyridine and o-xylene was not achieved.

[0040] The applicant declares that this invention illustrates the method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosols and its application through the above embodiments. However, this invention is not limited to the above embodiments, meaning that this invention does not necessarily rely on the above embodiments for implementation. Those skilled in the art should understand that any improvements to this invention, equivalent substitutions of raw materials for the product, addition of auxiliary components, and selection of specific methods all fall within the protection and disclosure scope of this invention.

[0041] The preferred embodiments of the present invention have been described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the technical concept of the present invention, various simple modifications can be made to the technical solution of the present invention, and these simple modifications all fall within the protection scope of the present invention.

[0042] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not describe the various possible combinations separately.

Claims

1. A method for detecting volatile and semi-volatile organic compounds in a heated cigarette aerosol, characterized by, The method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol includes the following steps: (1) The heated cigarette to be tested is inhaled, and the aerosol is captured using a filter and an adsorbent; (2) The filter containing the aerosol and the adsorbent are mixed with the internal standard and solvent for extraction to obtain the extract; (3) The extract was subjected to gas chromatography-tandem mass spectrometry for detection. The content of volatile and semi-volatile organic compounds in the heated cigarette to be tested was obtained based on the detection results and the standard curve. The volatile and semi-volatile organic compounds include any one or a combination of at least two of the following: vinyl chloride, 1,3-butadiene, isoprene, ethylene oxide, propylene oxide, benzene, acrylonitrile, toluene, ethylbenzene, p-xylene, m-xylene, o-xylene, pyridine, styrene, glycidyl ether, acetamide, nitrobenzene, acrylamide, or quinoline.

2. The method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol according to claim 1, characterized in that, The volatile and semi-volatile organic compounds include vinyl chloride, 1,3-butadiene, isoprene, ethylene oxide, propylene oxide, benzene, acrylonitrile, toluene, ethylbenzene, p-xylene, m-xylene, o-xylene, pyridine, styrene, glycidyl ether, acetamide, nitrobenzene, acrylamide, and quinoline.

3. The method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol according to claim 1 or 2, characterized in that, The internal standard in step (2) includes benzene-d6.

4. The method of detecting volatile, semi-volatile organic compounds in heated cigarette aerosol according to any one of claims 1 to 3, wherein, The solvent in step (2) is a carbon disulfide-methanol mixed solvent, wherein the volume ratio of carbon disulfide to methanol is 1:(3-5).

5. The method of detecting volatile, semi-volatile organic compounds in heated cigarette aerosol according to any one of claims 1-4, wherein, In step (3), the gas chromatography-tandem mass spectrometry column is an Agilent DB-WAX UI column.

6. The method of detecting volatile, semi-volatile organic compounds in heated cigarette aerosol according to any one of claims 1-5, wherein, In step (3), the temperature program for gas chromatography-tandem mass spectrometry is as follows: The initial temperature is 35-43℃, held for 4-6 minutes, then increased to 95-105℃ at 3-5℃ / min, then increased to 220-240℃ at 45-55℃ / min, held for 5-7 minutes, and then run for 2-4 minutes.

7. The method of detecting volatile, semi-volatile organic compounds in heated cigarette aerosol according to any one of claims 1 to 6, wherein, In step (3), the carrier gas flow rate in the gas chromatography-tandem mass spectrometry is 0.8-1.2 mL / min.

8. The method of detecting volatile, semi-volatile organic compounds in heated cigarette aerosol according to any one of claims 1-7, wherein, The standard curve in step (3) is prepared by a method comprising the following steps: A series of standard working solutions containing standard volatile and semi-volatile organic compounds were prepared and mixed with internal standards for gas chromatography-tandem mass spectrometry detection. A standard curve was constructed with the ratio of the quantitative ion peak area of ​​volatile and semi-volatile organic compounds in each standard working solution to that of the internal standard as the ordinate and the content of volatile and semi-volatile organic compounds in each standard working solution as the abscissa.

9. The application of the method for detecting volatile and semi-volatile organic compounds in heated cigarette aerosol according to any one of claims 1-8 in the analysis of harmful components in heated cigarettes.

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