CN111307961A - Simultaneous determination of 10 sour components in mainstream cigarette smoke - Google Patents

Abstract

The invention discloses a method for simultaneously measuring 10 acid aroma components in main stream smoke of cigarettes. Extracting particulate matters of main stream smoke of cigarettes by using an extracting agent, derivatizing and heating supernate by using a derivatizing reagent, detecting a smoke sample by using gas chromatography-tandem mass spectrometry (GC-MS/MS), and simultaneously detecting the contents of 10 acid aroma substances such as formic acid, acetic acid, isovaleric acid, n-valeric acid, 3-methyl-2-butenoic acid, 3-methylvaleric acid, 4-methylvaleric acid, n-hexanoic acid, n-heptanoic acid, benzoic acid and the like by using an internal standard quantitative method. The method focuses on the component with the unique sensory characteristic of acid aroma in the cigarette smoke, eliminates the interference of non-acid aroma organic acid components, has the advantages of quick detection, high sensitivity, good selectivity, high accuracy and the like, and can effectively reflect the composition condition of the main material basis of the acid aroma characteristic of the smoke of different cigarette products.

Description

Simultaneous determination of 10 sour components in mainstream cigarette smoke

technical field

The invention relates to the technical field of aroma component detection, in particular to a method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke.

Background technique

Sour aroma is an aroma characteristic that exists widely in nature. Many fruits, vegetables, pastries and brewed foods include sour aroma in the flavor characteristics. Sourness perception is also included in the complex flavor profile provided by cigarette smoke. Appropriate sour aroma can enhance the flavor quality of the above system and even bring unforgettable characteristics, while excessive sour aroma often leads to sensory impressions such as spoilage and sour odor, which reduces the acceptance of the product. Therefore, it is of great significance to rationally evaluate and monitor the basic composition of acid and aroma characteristic substances in the flavor system.

Although sour components are basically organic acids, not all organic acids are sour components. This point has not received enough attention in many reports. In fact, molecules that can interact with olfactory receptors to generate aromas themselves need to have a relatively low molecular weight, and all aroma substances found so far have molecular weights below 350 [Chemistry & Biodiversity, 2004, 1(12): 1957-1974], so the molecular weight Organic acids greater than 350 have no sour aroma contribution. Secondly, from an evolutionary point of view, the normal height of the human nasal cavity is 1.5-2 meters above the ground. The behavioral pattern of human smell determines that the human olfactory system is mainly used to detect volatile substances; in contrast, wild boars or hounds, etc. Animals can inhale non-volatile chemicals into the nasal cavity by holding their noses close to the ground and inhaling quickly, thus evolving the ability to detect low-volatile molecules [Predator-Prey Dynamics: The Role of Olfaction. New York, CRCPress, 2007] , so the less volatile organic acids have basically no acid flavor contribution. Third, there is no effective theoretical model to predict the relationship between aroma characteristics and chemical structure [Angew.Chem.Int.Ed., 2006, 45:6254–6261], and volatile small-molecule organic acids are often not acid aroma components For example, long-chain fatty acids such as oleic acid and linoleic acid are mainly oily, cinnamic acid is mainly spicy and balsamic, 2-ketobutyric acid is mainly pyrosweet, and crotonic acid is mainly milky. The research object of the analysis method is to focus on the organic acid component groups with real acid aroma characteristics, which is an important step to improve the effect of the material basis analysis of acid aroma characteristics.

At present, there are few quantitative analysis methods specifically aimed at the acid aroma components in mainstream cigarette smoke. Most of the methods use various organic acids with different aroma or taste characteristics in the smoke as the analysis object. Therefore, it is necessary to develop a rapid, sensitive and focused quantitative analysis method for important acid and aroma components in cigarette smoke.

SUMMARY OF THE INVENTION

The object of the present invention is to overcome the deficiencies of the prior art, and to provide a method for simultaneously measuring 10 kinds of sour aroma components in mainstream cigarette smoke. The method has the advantages of rapid detection, high sensitivity, good selectivity and high accuracy, and is suitable for For the determination of the content of sour aroma components in cigarette smoke.

In order to solve the above-mentioned technical problems, the concrete technical scheme adopted in the present invention is:

S1. Sample extraction and derivatization:

After capturing the mainstream cigarette smoke particulate matter with the Cambridge filter, transfer the Cambridge filter into the extraction container, add dichloromethane solution, and then add the internal standard solution for shaking extraction, take the supernatant and add the derivatization reagent to heat after heating. get flue gas samples;

The internal standard is trans-3-hexenoic acid;

S2. Sample gas chromatography-tandem mass spectrometry detection:

The flue gas samples were detected by GC-MS/MS, and the contents of 10 acid and aroma components were detected by the internal standard quantitative method;

The gas chromatography conditions for the detection are as follows: chromatographic column: DB-5MS (60m×0.25mm×0.25μm); carrier gas: He; column flow rate: 1 mL/min; inlet temperature: 250°C; temperature program: 50°C (1min), 5°C/min→200°C (0min); split ratio: 5:1.

In the present invention, the sour aroma components in the mainstream smoke of cigarettes are formic acid, acetic acid, isovaleric acid, n-valeric acid, 3-methyl-2-butenoic acid, 3-methylvaleric acid, 4-methyl valeric acid valeric acid, n-hexanoic acid, n-heptanoic acid, benzoic acid.

Preferably, the amount of the internal standard solution is 10-100 μL, the amount of the dichloromethane is 10-50 mL, and the derivatization reagent is N,O-bis(trimethylsilyl)trifluoroacetamide Or N-methyl-N-(trimethylsilane) trifluoroacetamide, and the amount of the derivatization reagent is 50-100 μL.

Preferably, in S1., the concentration of the internal standard is 2 mg/mL.

Preferably, the shaking extraction time is 20-50 min.

Preferably, in S1., the heating temperature is 20-100° C., and the heating time is 50-100 min.

Preferably, the mass spectrometry conditions for detection described in S2. are: GC/MS transmission line temperature: 250°C, EI ion source temperature: 230°C, quadrupole temperature: 150°C; EI ionization energy: 70eV; scanning mode: selected ion Scan, divide the time period according to the retention time.

Preferably, in S2., the standard working solution used in the internal standard quantification method contains 10 kinds of acid and aroma components and 1 kind of internal standard standard substance, and the solvent is dichloromethane; the internal standard substance in all standard working solutions is trans- The concentration of 3-hexenoic acid is 2 mg/mL, the concentration order of formic acid in the standard working solution is 0.645, 1.29, 3.225, 6.45, 12.9 μg/mL from low to high; the concentration order of acetic acid is from low to high 4.708, 9.416, 23.54, 47.08, 94.16 μg/mL; the concentration order of isovaleric acid from low to high is 0.187, 0.374, 0.935, 1.87, 3.74 μg/mL; the concentration order of n-valeric acid from low to high The highest concentrations were 0.051, 0.102, 0.255, 0.51, 1.02 μg/mL; the concentrations of 3-methyl-2-butenoic acid from low to high were 0.011, 0.022, 0.055, 0.11, 0.22 μg/mL; 3 -The concentration order of methylvaleric acid from low to high is 0.029, 0.058, 0.145, 0.29, 0.58μg/mL; 0.47, 0.94 μg/mL; the concentration order of n-hexanoic acid from low to high is 0.05, 0.1, 0.25, 0.5, 1 μg/mL; the concentration order of n-heptanoic acid from low to high is 0.094, 0.188, 0.47, 0.94, 1.88 μg/mL; the order of benzoic acid concentration from low to high was 0.94, 1.88, 4.7, 9.4, 18.8 μg/mL.

Compared with the prior art, the present invention has the following beneficial effects:

The invention fills in the blank of the prior art, and adopts the GC-MS/MS method to simultaneously measure the contents of 10 kinds of sour aroma components in mainstream cigarette smoke, specifically including formic acid, acetic acid, isovaleric acid, n-valeric acid, 3-methyl- 2-Butenoic acid, 3-methylvaleric acid, 4-methylvaleric acid, n-hexanoic acid, n-heptanoic acid, benzoic acid. Among them, the content of 3-methyl-2-butenoic acid and other components can be quantitatively detected in tobacco products for the first time. The method has the advantages of focusing on important sour components in smoke, rapid detection, high sensitivity, good selectivity and high accuracy, and is suitable for determining the content of sour components in cigarette smoke.

Description of drawings

Figure 1 shows the SIM images obtained by GC-MS detection of 10 standard samples of acid and aroma components.

FIG. 2 is a SIM diagram of the sour aroma components in the mainstream cigarette smoke sample of Example 1 detected by GC-MS.

Among them, 1: formic acid; 2: acetic acid; 3: isovaleric acid; 4: n-valeric acid; 5: 3-methyl-2-butenoic acid; 6: 3-methylvaleric acid; 7: 4-methyl valeric acid; 8: n-hexanoic acid; 9: n-heptanoic acid; 10: benzoic acid.

FIG. 3 is a schematic diagram of the whole process of the method of the present invention.

Detailed ways

The present invention will be further described below in conjunction with specific embodiments.

The present invention adopts the standard addition method to compare the retention time, and then quantifies by the internal standard curve method. The names and quantitative ion information of the 10 acid-flavor components and internal standards are shown in Table 1:

Table 1 10 acid-flavor components and their internal standard names and quantitative ion information

Under the detection conditions of the present invention, the quantitative curves, linear ranges, detection limits and quantification limits of the 10 acid-flavor components are shown in Table 2.

Table 2 Quantitative curve, linear range, detection limit and quantification limit of 10 acid-flavor components

Example 1

The method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke of the present embodiment includes the following steps:

S1. Puff the balanced and screened cigarettes of brand 1 according to the standard smoking conditions specified in YC/T29-1996, and use two Cambridge filters to capture the mainstream smoke particulate matter of 5 cigarettes, and place them in 50 mL In a stoppered conical flask, add 25 mL of dichloromethane solution, shake on a shaker for 20 min at room temperature, add 30 μL of trans-3-hexenoic acid internal standard solution, mix well, take 1.5 mL of supernatant, and add derivatization reagent 60 μL of N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) was placed in a 60°C water bath and heated for 50min to obtain a cigarette smoke sample.

S2. Use GC-MS to detect the flue gas sample obtained in step S1

The chromatographic conditions are: column: DB-5MS (60m×0.25mm×0.25μm); carrier gas: He; column flow rate: 1mL/min; ℃/min→200℃(0min); split ratio: 5:1.

The mass spectrometry conditions were: GC/MS transfer line temperature: 250 °C, EI ion source temperature: 230 °C, quadrupole temperature: 150 °C; EI ionization energy: 70 eV; scanning mode: selective ion scanning, and time periods were divided according to retention time.

Table 3 shows the test results of the cigarette of brand 1 by the method for simultaneously measuring 10 kinds of sour aroma components in mainstream cigarette smoke in this embodiment.

Example 2

The method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke of the present embodiment includes the following steps:

S1. Puff the balanced and screened cigarettes of brand 2 according to the standard smoking conditions specified in YC/T29-1996, and use two Cambridge filters to capture the mainstream smoke particles of 8 cigarettes, and place them in 50 mL In a stoppered conical flask, add 40 mL of dichloromethane solution, shake on a shaker for 30 min at room temperature, add 20 μL of trans-3-hexenoic acid internal standard solution, mix well, take 1 mL of supernatant, and add derivatization reagent BSTFA 70μL, placed in a 60°C water bath for 70min to obtain a cigarette smoke sample.

S2. use GC-MS to detect the flue gas sample obtained in step S1;

The chromatographic conditions are: column: DB-5MS (60m×0.25mm×0.25μm); carrier gas: He; column flow rate: 1mL/min; ℃/min→200℃(0min); split ratio: 5:1.

The mass spectrometry conditions were: GC/MS transfer line temperature: 250 °C, EI ion source temperature: 230 °C, quadrupole temperature: 150 °C; EI ionization energy: 70 eV; scanning mode: selective ion scanning, and time periods were divided according to retention time.

Table 3 shows the test results of the cigarettes of brand 2 by the method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke in this embodiment.

Example 3

The method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke of the present embodiment includes the following steps:

S1. Puff the balanced and screened cigarettes of brand 3 according to the standard smoking conditions specified in YC/T29-1996, and use two Cambridge filters to capture the mainstream smoke particulate matter of 8 cigarettes, and place them in 50 mL In a stoppered conical flask, add 50 mL of dichloromethane solution, shake on a shaker for 40 min at room temperature, add 60 μL of trans-3-hexenoic acid internal standard solution, mix well, take 1 mL of supernatant, and add derivatization reagent BSTFA 90 μL, placed in a 50°C water bath and heated for 90 min to obtain a cigarette smoke sample.

S2. use GC-MS to detect the flue gas sample obtained in step S1;

The spectral conditions are: chromatographic column: DB-5MS (60m×0.25mm×0.25μm); carrier gas: He; column flow rate: 1mL/min; ℃/min→200℃(0min); split ratio: 5:1.

The mass spectrometry conditions were: GC/MS transfer line temperature: 250 °C, EI ion source temperature: 230 °C, quadrupole temperature: 150 °C; EI ionization energy: 70 eV; scanning mode: selective ion scanning, and time periods were divided according to retention time.

Table 3 shows the test results of the cigarettes of brand 3 by the method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke in this embodiment.

Example 4

The method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke of the present embodiment includes the following steps:

S1. Puff the balanced and screened cigarettes of brand 4 according to the standard smoking conditions specified in YC/T29-1996, and use two Cambridge filters to capture the mainstream smoke particulate matter of 8 cigarettes, and place them in 50 mL In a stoppered conical flask, add 50 mL of dichloromethane solution, shake on a shaker for 30 min at room temperature, add 100 μL of trans-3-hexenoic acid internal standard solution, mix well, take 1 mL of supernatant, and add 100 μL of derivatization reagent BSTFA , placed in a 60 ℃ water bath and heated for 100 min to obtain cigarette smoke samples.

S2. use GC-MS to detect the flue gas sample obtained in step S1;

The spectral conditions are: chromatographic column: DB-5MS (60m×0.25mm×0.25μm); carrier gas: He; column flow rate: 1mL/min; ℃/min→200℃(0min); split ratio: 5:1.

The mass spectrometry conditions were: GC/MS transfer line temperature: 250 °C, EI ion source temperature: 230 °C, quadrupole temperature: 150 °C; EI ionization energy: 70 eV; scanning mode: selective ion scanning, and time periods were divided according to retention time.

Table 3 shows the test results of cigarettes of brand 4 by the method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke in this embodiment.

Example 5

The method for simultaneously measuring 10 sour aroma components in mainstream cigarette smoke of the present embodiment includes the following steps:

S1. Puff the balanced and screened cigarettes of brand 5 according to the standard smoking conditions specified in YC/T29-1996, and use two Cambridge filters to capture the mainstream smoke particulate matter of 8 cigarettes, and place them in 50 mL In a stoppered conical flask, add 35 mL of dichloromethane solution, shake on a shaker for 40 min at room temperature, add 70 μL of trans-3-hexenoic acid internal standard solution, mix well, take 1 mL of supernatant, and add derivatization reagent BSTFA 80μL, placed in a water bath at 70°C for 60min, to obtain a cigarette smoke sample.

S2. use GC-MS to detect the flue gas sample obtained in step S1;

The spectral conditions are: chromatographic column: DB-5MS (60m×0.25mm×0.25μm); carrier gas: He; column flow rate: 1mL/min; ℃/min→200℃(0min); split ratio: 5:1.

The mass spectrometry conditions were: GC/MS transfer line temperature: 250 °C, EI ion source temperature: 230 °C, quadrupole temperature: 150 °C; EI ionization energy: 70 eV; scanning mode: selective ion scanning, and time periods were divided according to retention time.

Table 3 shows the test results of the cigarette of brand 5 by the method for simultaneously measuring 10 kinds of sour aroma components in mainstream cigarette smoke in this embodiment.

The detection result of acid aroma component content in the cigarette samples of table 3 embodiment 1～5

The equilibrated and screened cigarettes were smoked according to the standard smoking conditions specified in YC/T29-1996, and 8 cigarette smoke particles were captured with 2 Cambridge filters. Measured according to the method of Example 1, 6 groups were measured in parallel, and the RSD values of the 6 parallel measured values of 10 sour aroma components were between 3% and 8% (see Table 4), which proves that the method of the present invention is more accurate than high and repeatable.

Table 4 Precision of solvent extraction-gas chromatography-mass spectrometry method

Obviously, the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those of ordinary skill in the art, changes or modifications in other different forms can also be made on the basis of the above description. There is no need and cannot be exhaustive of all implementations here. Any modifications, equivalent replacements and improvements made within the spirit and principle of the present invention shall be included within the protection scope of the claims of the present invention.

Claims (8)

1. A method for simultaneously measuring the contents of 10 acid aroma components in the mainstream smoke of cigarettes is characterized by comprising the following steps: the method comprises the following steps:

s1, sample extraction and derivatization:

after particulate matters of mainstream smoke of the cigarettes are captured by using a Cambridge filter, transferring the Cambridge filter into an extraction container, adding a dichloromethane solution, adding an internal standard substance solution for oscillation extraction, taking supernate, adding a derivatization reagent, and heating to obtain a smoke sample;

the internal standard substance is trans-3-hexenoic acid;

s2, detecting a sample by gas chromatography-tandem mass spectrometry:

detecting a smoke sample by adopting GC-MS/MS, and detecting the content of 10 acid and fragrance components by an internal standard quantitative method;

the gas chromatography conditions for detection are as follows: a chromatographic column: DB-5MS (60m 0.25mm 0.25 μm); carrier gas: he; column flow rate: 1 mL/min; sample inlet temperature: 250 ℃; temperature programming: 50 deg.C (1min),5 deg.C/min → 200 deg.C (0 min); the split ratio is as follows: 5:1.

2. The method for simultaneously measuring 10 acid aroma components in the mainstream smoke of the cigarette according to claim 1, wherein the acid aroma components in the mainstream smoke of the cigarette are formic acid, acetic acid, isovaleric acid, n-valeric acid, 3-methyl-2-butenoic acid, 3-methylvaleric acid, 4-methylvaleric acid, n-hexanoic acid, n-heptanoic acid and benzoic acid.

3. The method for simultaneously determining 10 acid aroma components in mainstream smoke of cigarettes according to claim 1, wherein the amount of the internal standard substance solution is 10-100 μ L, the amount of dichloromethane is 10-50 mL, the derivatization reagent is N, O-bis (trimethylsilyl) trifluoroacetamide or N-methyl-N- (trimethylsilane) trifluoroacetamide, and the amount of the derivatization reagent is 50-100 μ L.

4. The method for simultaneously determining 10 acid-flavor components in mainstream smoke of cigarettes according to claim 1, wherein the method comprises the following steps: s1, the concentration of the internal standard substance is 2 mg/mL.

5. The method for simultaneously measuring 10 acid aroma components in the mainstream smoke of cigarettes according to claim 1, wherein in S1, the shaking extraction time is 20-50 min.

6. The method for simultaneously measuring 10 acid aroma components in the mainstream smoke of the cigarettes according to claim 1, wherein in S1, the heating temperature is 20-100 ℃, and the heating time is 50-100 min.

7. The method for simultaneously determining 10 acid-flavor components in mainstream smoke of cigarettes according to claim 1, wherein the method comprises the following steps: s2, the mass spectrum conditions of the detection are as follows: GC/MS transmission line temperature: 250 ℃, EI ion source temperature: 230 ℃, quadrupole temperature: 150 ℃; EI ionization energy: 70 eV; scanning mode: an ion scan is selected, and the time segments are divided according to retention time.

8. The method for simultaneously determining 10 acid-flavor components in mainstream smoke of cigarettes according to claim 1, wherein in S2, the standard working solution adopted by the internal standard quantitative method contains 10 acid-flavor components and 1 internal standard substance, and the solvent is dichloromethane; the concentration of the internal standard substance trans-3-hexenoic acid in all the standard working solutions is 2mg/mL, and the concentration levels of formic acid in the standard working solutions are 0.645, 1.29, 3.225, 6.45 and 12.9 mu g/mL from low to high in sequence; the concentration levels of the acetic acid are 4.708, 9.416, 23.54, 47.08 and 94.16 mu g/mL from low to high in sequence; the concentration magnitude of the isovaleric acid is 0.187, 0.374, 0.935, 1.87 and 3.74 mu g/mL from low to high in sequence; the concentration magnitude of the n-valeric acid is 0.051, 0.102, 0.255, 0.51 and 1.02 mu g/mL from low to high in sequence; the concentration levels of the 3-methyl-2-butenoic acid are 0.011, 0.022, 0.055, 0.11 and 0.22 mu g/mL from low to high in sequence; the concentration levels of the 3-methylvaleric acid are 0.029, 0.058, 0.145, 0.29 and 0.58 mu g/mL from low to high in sequence; the concentration levels of the 4-methylvaleric acid are 0.047, 0.094, 0.235, 0.47 and 0.94 mu g/mL from low to high in sequence; the concentration magnitude of the n-hexanoic acid is 0.05, 0.1, 0.25, 0.5 and 1 mu g/mL from low to high in sequence; the concentration levels of the n-heptanoic acid are 0.094, 0.188, 0.47, 0.94 and 1.88 mu g/mL from low to high in sequence; the concentration levels of the benzoic acid are 0.94 mu g/mL, 1.88 mu g/mL, 4.7 mu g/mL, 9.4 mu g/mL and 18.8 mu g/mL from low to high.

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