Method for detecting per-puff dissolution amount of characteristic components of acid-sweet flavored cigarette tipping paper and method for studying per-puff dissolution regularity

Abstract

The application provides a method for detecting the mouth-by-mouth dissolution amount of characteristic components of acid-sweet flavor cigarette tipping paper and a method for researching the mouth-by-mouth dissolution rule. The method simulates the mouth-by-mouth dissolution of the flavor tipping paper cigarette to obtain a mouth-by-mouth dissolution sample solution meeting the quantitative analysis requirement; meanwhile, the liquid chromatography-tandem mass spectrometry method is used to simultaneously detect the acid-sweet characteristic components; the acid-sweet characteristic components include acidifiers: citric acid and malic acid; sweeteners: acesulfame potassium, sodium saccharin and sucralose. The application first realizes the quantitative detection of the mouth-by-mouth dissolution amount of the acid-sweet flavor cigarette tipping paper characteristic components, improves the detection efficiency, realizes the mouth-by-mouth dissolution rule research of the acid-sweet flavor cigarette tipping paper characteristic components, and provides technical support for the quality monitoring in the production process of the flavor cigarette tipping paper.

Description

Technical Field

[0001] This invention belongs to the field of chemical component detection in cigarette tipping paper, specifically involving a method for detecting the leaching amount of characteristic components of sweet and sour flavored cigarette tipping paper per puff and a method for studying the leaching pattern per puff. Background Technology

[0002] With the tobacco industry's continuous increasing demands for product innovation, the research and development of tipping paper for sweet and sour flavored cigarettes has become a hot topic in the industry. The puff-by-puff release stability of sweet and sour flavored cigarette tipping paper directly affects consumers' sensory experience and is a key factor in measuring product quality consistency. Therefore, systematically studying the puff-by-puff dissolution patterns of characteristic components on sweet and sour flavored cigarette tipping paper is of great significance for product development and quality control. Quantitative detection of the characteristic components of acidulants and sweeteners on tipping paper is an important and necessary step in studying the puff-by-puff dissolution patterns of characteristic components on sweet and sour flavored cigarette tipping paper. However, the composition of tipping paper is complex, and the content of the characteristic components of acidulants and sweeteners in it is at the trace level, which increases the difficulty of simultaneous quantitative detection of acidulants and sweeteners on tipping paper.

[0003] Currently, the analysis of characteristic components of sweeteners such as acesulfame potassium, sodium saccharin, and sucralose in cigarette tipping paper mainly employs techniques such as high-performance liquid chromatography-evaporative light scattering detection (HPLC-ELSD) and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS / MS). While no methods have been reported in the literature for detecting acidulants such as citric acid and malic acid in tipping paper, mature analytical methods exist for related matrices in food and tobacco. Due to the high polarity of acidulants, they are typically determined using methyl esterification derivatization-gas chromatography-flame ionization detector (GC-FID), or ion chromatography (IC) and high-performance liquid chromatography (HPLC). The matrix significantly influences the quantitative detection methods for acidulants and sweeteners; different matrices may lead to different detection methods for the same acidulants and sweeteners. Therefore, studying the dissolution patterns of sweeteners and acidulants in sweet and sour flavored cigarette tipping paper separately would require two independent sets of instruments, resulting in cumbersome experimental procedures and long detection cycles. Meanwhile, when using the existing methods described above to analyze the dissolution amounts of acidulants and sweeteners in tipping paper, a large number of tipping paper samples are generally required for extraction in order to achieve quantitative analysis, which makes sample pretreatment cumbersome and time-consuming. Summary of the Invention

[0004] In view of this, in order to solve the problem of puff-by-puff dissolution and detection of acidulants and sweeteners on sweet and sour flavored cigarette tipping paper, and further study their puff-by-puff dissolution rules, this invention uses artificial lips to simulate puff-by-puff dissolution of flavored cigarette tipping paper. In order to meet the requirements of quantitative analysis, a multi-puff accumulation method is adopted. The dissolution solution is detected by HPLC-MS / MS. This invention successfully realizes the detection of the puff-by-puff dissolution amount and the study of the puff-by-puff dissolution rules of the characteristic components of the two types of additives in sweet and sour flavored cigarette tipping paper, thereby simplifying the sample pretreatment method and significantly improving the efficiency of analysis.

[0005] Specifically, the technical solution adopted in this invention is as follows: This invention provides a method for detecting the leaching amount of characteristic components of sweet and sour flavored cigarette tipping paper in each puff, comprising the following steps: Sample pretreatment: Simulate the puff-by-puff dissolution of sweet and sour flavored cigarettes to obtain multiple puff-by-puff dissolution samples to be tested: wherein, the characteristic components of the tipping paper for the sweet and sour flavored cigarettes include acidulants and sweeteners, the acidulants include citric acid and malic acid, and the sweeteners include acesulfame potassium, sodium saccharin and sucralose. HPLC-MS / MS detection: The acidulant and sweetener in each of the test slurry samples were simultaneously and quantitatively detected using the HPLC-MS / MS method, wherein the HPLC-MS / MS method included the following liquid chromatography detection conditions: Liquid chromatography column: InfinityLab Poroshell 120 SB C 18 ; The mobile phase consists of an aqueous solution of ammonium acetate and methanol in a volume ratio of (0.8–10): (0–9.2), wherein the ammonium acetate in the aqueous solution is an additive. Elution method: gradient elution.

[0006] Furthermore, the sample pretreatment step includes: using an artificial lip simulation device to simulate a smoker taking puffs of multiple sweet and sour flavored cigarettes, accumulating the dissolution liquid corresponding to the number of puffs when the multiple sweet and sour flavored cigarettes are taken puff by puff, and obtaining multiple samples of the puff-by-puff dissolution liquid to be tested.

[0007] Furthermore, the sample pretreatment steps include: S1. Provides multiple artificial lip simulation devices, and each artificial lip simulation device includes a container and a sponge filled in the container and soaked in artificial saliva, wherein the sponge has a cigarette insertion hole in the middle that matches the diameter of the sweet and sour flavored cigarette. S2. Insert the tip of a sweet and sour flavored cigarette into the cigarette insertion holes of multiple artificial lip simulation devices in sequence, so that the characteristic components of the tip of the sweet and sour flavored cigarette dissolve into the corresponding sponge, and take out the sweet and sour flavored cigarette from the corresponding sponge to complete the dissolution experiment for a single sweet and sour flavored cigarette corresponding to the number of puffs, so that multiple sponges corresponding to the number of puffs contain the tip of the corresponding number of puffs dissolving liquid; S3. Repeat step S2 using multiple sweet and sour flavored cigarettes, and accumulate the corresponding number of sponge paper leachate in the sponge with the corresponding number of puffs. S4. Using water as the extraction solvent, extract the sponge containing the sponge with the corresponding number of suction ports to obtain multiple samples of the sponge to be tested.

[0008] Furthermore, in step S4, an internal standard composition is added to the extraction solvent, and the internal standard composition includes... 13 C6-citric acid and sucralose-d6, in the sample of the dissolution solution to be tested 13 The concentrations of C6-citric acid and sucralose-d6 were both 5–10 ng / mL.

[0009] Furthermore, the HPLC-MS / MS method includes the following mass spectrometry detection conditions: electrospray ionization source, negative ion scanning mode; detection mode: multiple reaction monitoring.

[0010] This invention also provides a method for studying the dissolution pattern of characteristic components of sweet and sour flavored cigarette tipping paper, comprising the following steps: Sample pretreatment: Simulate the puff-by-puff dissolution of sweet and sour flavored cigarettes to obtain multiple puff-by-puff dissolution samples to be tested: wherein, the characteristic components of the tipping paper for the sweet and sour flavored cigarettes include acidulants and sweeteners, the acidulants include citric acid and malic acid, and the sweeteners include acesulfame potassium, sodium saccharin and sucralose. HPLC-MS / MS detection: The acidulant and sweetener in each of the sip-by-sip dissolution samples were simultaneously quantitatively detected using the HPLC-MS / MS method to obtain the sip-by-sip dissolution amount of the acidulant and sweetener; wherein the HPLC-MS / MS method includes the following liquid chromatography detection conditions: Liquid chromatography column: InfinityLab Poroshell 120 SB C 18 ; The mobile phase consists of an aqueous solution of ammonium acetate and methanol in a volume ratio of (0.8–10): (0–9.2), wherein the ammonium acetate in the aqueous solution is an additive. Elution method: gradient elution; Regularity study: The dissolution rate of the acidulants and sweeteners was studied and analyzed to obtain the dissolution rate regularity of characteristic components of sweet and sour flavored cigarette tipping paper.

[0011] Therefore, the technical solution provided by this invention obtains a sample solution that meets the requirements of quantitative analysis by simulating the dissolution of flavored cigarette tipping paper in each puff. Simultaneously, through screening of liquid chromatography columns and optimization of gradient elution conditions, on the one hand, a systematic screening and evaluation of a large number of liquid chromatography columns is conducted to select columns capable of separating five characteristic components: citric acid, malic acid, acesulfame potassium, sodium saccharin, sucralose, and acesulfame potassium. On the other hand, the composition of the mobile phase and gradient elution conditions are optimized, thus achieving effective separation of the above five characteristic components from both acidulants and sweeteners. This enables simultaneous detection of the characteristic components of sweet and sour flavored cigarette tipping paper in each puff, improving detection efficiency and thus enabling research on the dissolution law of characteristic components of sweet and sour flavored cigarette tipping paper, providing technical support for quality monitoring during the production process of flavored cigarette tipping paper. Attached Figure Description

[0012] Figure 1 A schematic diagram of the artificial lip simulation device provided by the present invention; wherein, 1: container, 2: cigarette insertion hole, 3: sponge; Figure 2-4 The above are HPLC-MS / MS chromatograms of the characteristic component standards and internal standards in Example 1 of this invention. The characteristic components include citric acid, malic acid, acesulfame potassium, sodium saccharin, and sucralose standards. The internal standards include... 13 C6-citric acid and sucralose-d6; Figure 5 For Comparative Example 1, XBridge® C was used. 18 HPLC-MS / MS chromatogram of citric acid standard on a chromatographic column (5µm, 2.1×150mm); Figure 6 The HPLC-MS / MS chromatogram of sodium saccharin standard under the gradient elution conditions used in Comparative Example 4 is shown below. Figure 7 The HPLC-MS / MS chromatogram of the citric acid standard is shown when the mobile phase shown in Comparative Example 5 is used. Detailed Implementation

[0013] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0014] Unless otherwise specified, the terminology used in this invention is a common term in the relevant field. The technical means used in the embodiments, such as sample preparation methods, detection methods, detection conditions, etc., are all conventional means well known to those skilled in the art unless otherwise specified. The reagents and products used are also commercially available.

[0015] To address the current lack of a research method for studying the puff-by-puff dissolution patterns of characteristic components in sweet and sour flavored cigarette tipping paper, this invention employs a biomimetic artificial lip to simulate the puff-by-puff dissolution of these components during smoking. After the puff-by-puff dissolution is complete, the sweet and sour components on the artificial lip are extracted using a solvent and detected using HPLC-MS / MS. This allows for the study of the puff-by-puff dissolution patterns of characteristic components in sweet and sour flavored cigarette tipping paper. Specifically, on the one hand, the artificial lip achieves a biomimetic simulation of the puff-by-puff dissolution of sweet and sour components during smoking; on the other hand, the HPLC-MS / MS method enables the simultaneous detection of acidulants and sweeteners, thus significantly improving analytical efficiency. The specific implementation scheme of this invention is as follows: This invention provides a method for detecting the leaching amount of characteristic components of sweet and sour flavored cigarette tipping paper in each puff, comprising the following steps: Sample pretreatment: Artificial lips were used to simulate the puff-by-puff dissolution of tipping paper used in sweet and sour flavored cigarettes. After multiple puffs, several samples of the puff-by-puff dissolution solution were obtained for testing. The tipping paper contained acidulants and sweeteners; the acidulants included citric acid and malic acid, and the sweeteners included acesulfame potassium, sodium saccharin, and sucralose. This step specifically included the following steps: S1. Provide 6 to 10 artificial lip simulation devices, and each artificial lip simulation device is as follows: Figure 1 As shown, it includes a container 1 and a sponge 3 filled in the container 1 and soaked in artificial saliva. The sponge 3 has a cigarette insertion hole 2 in the middle, which is equivalent to the diameter of the sweet and sour flavored cigarette. Preferably, the number of artificial lip simulation devices is equal to the number of puffs that can be taken with the sweet and sour flavored cigarette. That is, 6 to 10 artificial lip simulation devices are used to simulate a smoker taking 6 to 10 puffs of the sweet and sour flavored cigarette. S2. Insert the tip of a sweet and sour flavored cigarette into the sponge of the first artificial lip simulation device to a depth of 1 cm, let it dissolve for 2 seconds, then remove it and let it stand for 30 seconds; then insert it into the sponge of the second artificial lip simulation device, let it dissolve for 2 seconds, remove it and let it stand for 30 seconds, then insert it into the sponge of the third artificial lip simulation device, and so on, until the sweet and sour flavored cigarette is finished smoking; so that 6 to 10 sponges corresponding to the number of puffs contain the tip of the dissolving liquid corresponding to the number of puffs. S3. Since the single-puff dissolution of characteristic components in the tipping paper of sweet and sour flavored cigarettes is low, in order to meet the requirements of instrument quantitative analysis, follow the above step S2 and take 5 to 10 sweet and sour flavored cigarettes to accumulate dissolution puff by puff, so as to accumulate the tipping paper dissolution liquid corresponding to the number of puffs in the sponge of the corresponding number of puffs. S4. After the dissolution test, remove the sponge and add it to a 50–10 mL centrifuge tube. Add 10–50 mL of internal standard aqueous solution, and sonicate for 10–60 minutes. Take an appropriate amount of supernatant and filter it through a 0.22 µm aqueous filter membrane to obtain multiple dissolution samples for subsequent HPLC-MS / MS analysis. The internal standard aqueous solution is composed of water, 13 It is composed of C6-citric acid and sucralose-d6, in which 13 C6-citric acid serves as an internal standard for citric acid and malic acid, while sucralose-d6 serves as an internal standard for sodium saccharin, sucralose, and acesulfame potassium. The sample to be tested contains... 13 The concentrations of C6-citric acid and sucralose-d6 were both 5–10 ng / mL.

[0016] HPLC-MS / MS detection: Each sample of the analyte or the mixed solution of standard samples of each characteristic component was filtered through a 0.22 μm syringe aqueous filter and then transferred to a 1–3 mL autosampler vial for HPLC-MS / MS analysis (ABSCI 5500 triple quadrupole mass spectrometer). The HPLC detection conditions included: Column: InfinityLab Poroshell 120 SB C, 2.7µm, 3.0×150mm. 18 ; The mobile phase consists of water and methanol in a volume ratio of 1:1 to 1:9, wherein the organic phase is methanol and the aqueous phase is an aqueous solution of ammonium acetate, with an ammonium acetate concentration of 2 to 10 mmol / L; preferably, the concentration of the ammonium acetate aqueous solution is 4 to 6 mmol / L. Column temperature: 25-45 ℃; flow rate: 0.1~0.5 mL / min; injection volume: 8~12 μL; elution method: gradient elution, and gradient elution conditions are shown in Table 1.

[0017] Table 1 Gradient elution conditions Time (min) methanol Ammonium acetate aqueous solution 0 8～25% 92～75% 3.5 10～40% 90～60% 10 80～100% 20～0% 15 80～100% 20～0% 15.1 75～95% 25～5% 20 10～90% 90～10% The mass spectrometry detection conditions are as follows: Electrospray ionization source was used, with negative ion scanning mode; detection method: multiple reaction monitoring; electrospray voltage: 4500V; auxiliary gas 1 pressure: 40 psi; auxiliary gas 2 pressure: 40 psi; ion source temperature: 400 ℃. The characteristic components of acidulants (citric acid, malic acid) and sweeteners (sodium saccharin, sucralose, and acesulfame potassium) were analyzed using negative ion scanning mode; cooling agents (WS-23, WS-3) were analyzed using positive ion scanning mode; and the mass spectrometry parameters of each compound are shown in Table 2.

[0018] Table 2 Mass Spectrometry Parameters of the Target Compound Characteristic components Scanning mode Retention time Mother ion Quantitative ions Qualitative ions DP(v) CE(v) Citric acid negative ions 2.766 191.0 110.9 86.8 -60 -15 <![CDATA[ 13 C6-Citrate negative ions 2.759 197.0 116.9 92.8 -60 -15 malic acid negative ions 2.786 132.9 114.9 71.0 -60 -15 Acesulfame K negative ions 4.415 162.0 82.0 162.0 -40 -18 Sodium saccharin negative ions 5.639 182.1 106.1 42.0 -60 -25 Sucralose negative ions 9.639 395.0 359.0 361.1 -120 -16 <![CDATA[Sucralose-d6]]> negative ions 8.928 401.0 365.0 367.0 -120 -16 The technical solution provided by this invention uses an artificial lip simulation device to biomimetically simulate the dissolution of sweet and sour characteristic components in the tipping paper during a smoker's cigarette smoking. The above detection method is beneficial for the simultaneous detection of sweet and sour characteristic components, improving detection efficiency. This is conducive to studying the dissolution law of sweet and sour characteristic components in flavored cigarette tipping paper and shortening the research cycle. Example 1

[0019] This embodiment provides an HPLC-MS / MS method for detecting characteristic components of sweet and sour flavored cigarette tipping paper, including the following steps: Preparation of standard solutions: Dissolve each characteristic component standard in water as the extraction solvent to prepare a characteristic component standard stock solution; mix the characteristic component standard stock solution and the corresponding internal standard solution uniformly to obtain a characteristic component standard mixed solution, wherein the concentration of each characteristic component is 0.1–100 ng / mL; wherein the characteristic components include citric acid, malic acid, acesulfame potassium, sodium saccharin, and sucralose. The internal standard solution consists of an internal standard and water, wherein the internal standard includes… 13 C6-citric acid and sucralose-d6, the characteristic component standard mixed solution 13 The concentrations of C6-citric acid and sucralose-d6 were both 5–25 ng / mL.

[0020] Liquid chromatography detection: The standard solutions of each characteristic component were filtered through a 0.22 μm syringe aqueous filter and then transferred to 2 mL autosampler vials for HPLC-MS / MS analysis. The liquid chromatography detection conditions included: column: 2.7 µm, 3.0 × 150 mm InfinityLab Poroshell 120 SB C 18 Mobile phase: composed of water and methanol in a volume ratio of 1:1 to 1:9, wherein the organic phase is methanol and the aqueous phase is an aqueous solution of ammonium acetate with a concentration of 5 mmol / L; Column temperature: 40 ℃; Flow rate: 0.2 mL / min; Injection volume: 10 μL; Elution method: gradient elution, and the gradient elution conditions are shown in Table 3.

[0021] Table 3 Gradient elution conditions for liquid chromatography Time (min) methanol Ammonium acetate aqueous solution 0 15% 85% 3.5 30% 70% 10 90% 10% 15 90% 10% 15.1 15% 85% 20 15% 85% Mass spectrometry detection: Electrospray ionization source, negative ion scanning mode; Detection method: multiple reaction monitoring; Electrospray voltage: 4500 V; Assist gas 1 pressure: 40 psi; Assist gas 2 pressure: 40 psi; Ion source temperature: 400 ℃. The mass spectrometry parameters of each compound are shown in Table 2.

[0022] The HPLC-MS / MS detection results for each characteristic component at a concentration of 100 ng / mL and an internal standard of 20 ng / mL are as follows: Figures 2-4 As shown, the standard samples of citric acid, malic acid, sodium saccharin, sucralose, and acesulfame potassium (left figure of each) have high peak shapes and high responses, which can meet the requirements of quantitative analysis. Figures 2-4 (Right figure) demonstrates that HPLC-MS / MS can quantitatively detect the content of citric acid, malic acid, sodium saccharin, sucralose, and acesulfame potassium in tipping paper.

[0023] In the process of liquid chromatography detection, the selection of chromatographic column, mobile phase composition, elution conditions and other factors have an important impact on the quantitative determination results of five characteristic components involved in two types of additives in the characteristic components of sweet and sour flavored cigarette tipping paper: citric acid, malic acid, acesulfame potassium, sodium saccharin, sucralose and acesulfame potassium. Comparative Examples 1-3

[0024] Comparative Examples 1-3 each provide an HPLC-MS / MS method for detecting characteristic components of sweet and sour flavored cigarette tipping paper, which is basically the same as the detection method provided in Example 1, with the main difference being the use of different chromatographic columns.

[0025] Comparative Example 1: XBridge® C with a specification of 5µm and 2.1×150mm. 18 Chromatographic column; Comparative Example 2: A ZORBAX StablbBond C18 column with a specification of 5µm and a diameter of 2.1×150mm; Comparative Example 3: A Waters T3 column with a specification of 5µm and a size of 2.1×150mm.

[0026] from Figure 5 As can be seen from the data: when separating the seven characteristic components in the packaged paper using standard samples, Comparative Example 1 uses XBridge® C 18 The citric acid peak shape was poor and severe tailing occurred in the chromatographic column; Comparative Example 2 used a ZORBAX Stablb Bond C18 column and Comparative Example 3 used a Waters T3 column, and the citric acid peak shape was poor and severe tailing occurred in both. Therefore, with Figure 2 In comparison, it uses the Infinity Lab Poroshell 120 SB C 18 Other liquid chromatography columns with similar theoretical plate numbers may present difficulties in separating characteristic components, making it challenging to meet the requirements for quantitative analysis. Therefore, the Infinity Lab Poroshell 120 SB C is the preferred choice. 18 The chromatographic column is used for the analysis of multiple characteristic components in the tipping paper of cigarettes with a complex flavor based on sweetness. Comparative Example 4-5

[0027] Comparative Examples 4-5 each provide an HPLC-MS / MS method for detecting characteristic components of sweet and sour flavored cigarette tipping paper, which is basically the same as the detection method provided in Example 1. The main difference is that the liquid chromatography gradient conditions used in Comparative Examples 4-5 are different. Specifically, Comparative Example 4 uses the gradient elution conditions shown in Table 4, and the detection results are as follows: Figure 6 As shown in Table 5; Comparative Example 5 uses the gradient elution conditions shown in Table 5.

[0028] Table 4. Liquid chromatography gradient elution conditions used in Comparative Example 4 Time (min) methanol Ammonium acetate aqueous solution 0 2% 98% 3.5 2% 98% 10 90% 10% 15 90% 10% 15.1 2% 98% 20 2% 98% Table 5. Liquid chromatography gradient elution conditions used in Comparative Example 5 Time (min) methanol Ammonium acetate aqueous solution 0 15% 85% 3.5 30% 70% 10 90% 10% 15 90% 10% 20 15% 85% contrast Figure 3 and Figure 6 It can be seen that when the gradient elution conditions in Example 1 are used, the sodium saccharin peak is symmetrical, with a peak width of approximately 0.6 minutes. When the gradient elution conditions in Comparative Example 4 are used, the sodium saccharin peak is significantly broadened, with a peak width of approximately 2.5 minutes, which will significantly reduce the detection sensitivity of sodium saccharin. When the gradient elution conditions in Example 1 are used, the chromatographic peaks of each characteristic component are symmetrical, with peak widths all less than 1 minute. When the gradient elution conditions in Comparative Example 5 are used, the chromatographic peaks of each characteristic component are significantly broadened, with peak widths of approximately 2-3 minutes, which will significantly reduce the detection sensitivity of each target compound. Therefore, the liquid chromatography gradient elution conditions in Example 1 are the preferred gradient elution conditions for the HPLC-MS / MS analysis of characteristic components in sweet and sour flavor cigarette tipping paper. Comparative Example 6

[0029] Comparative Example 6 provides an HPLC-MS / MS method for detecting characteristic components of sweet and sour flavored cigarette tipping paper, which is basically the same as the detection method provided in Example 1, with the main difference being: the mobile phase in Comparative Example 6 is different; the gradient elution conditions are the same as in Table 3. Comparative Example 6 uses formic acid as the mobile phase additive, i.e., mobile phase A: methanol; B: 0.1 wt% formic acid aqueous solution; the detection results are as follows: Figure 7As shown.

[0030] Comparison Figure 2 and Figure 7 It can be seen that when 5 mmol / L ammonium acetate was added to mobile phase B in Example 1, the citric acid peak shape was good, meeting the requirements for quantitative analysis. However, when mobile phase B in Comparative Example 6 was 0.1 wt% formic acid aqueous solution, the citric acid peak shape deteriorated, with severe tailing, seriously affecting the accuracy of quantification. Furthermore, when mobile phase B was 0.1 wt% acetic acid aqueous solution; or when mobile phase A was 0.1 wt% ammonium acetate aqueous solution and mobile phase B was 0.1 wt% acetic acid aqueous solution, the citric acid peak shape deteriorated, affecting the accuracy of quantification. Therefore, in Example 1, the preferred chromatographic conditions for HPLC-MS / MS analysis of characteristic components in sweet and sour flavor cigarette tipping paper were mobile phase A: methanol and mobile phase B: 5 mmol / L ammonium acetate aqueous solution.

[0031] In summary, the HPLC-MS / MS detection conditions described in Example 1 above are beneficial for the liquid chromatographic separation of citric acid, malic acid, acesulfame potassium, sodium saccharin, sucralose, and acesulfame potassium. Combined with the accurate qualitative analysis of mass spectrometry, the accuracy of quantitative detection of characteristic components of sweeteners and acidulants in tipping paper can be improved. Example 2

[0032] This embodiment discloses a method for detecting the puff-by-puff dissolution amount of characteristic components in sweet and sour flavored cigarette tipping paper and a method for studying the puff-by-puff dissolution pattern. HPLC-MS / MS is used to quantitatively detect the puff-by-puff dissolution amount of characteristic components in sweet and sour flavored cigarette tipping paper 1. The specific detection method includes: Sample pretreatment: Figure 1 The artificial lip simulation device shown uses a sponge (2.8 cm in diameter) soaked in artificial saliva (5 mL) to simulate artificial lips (the sponge has a hole in the center with a diameter similar to that of a cigarette). The sponge is placed in a glass container (3 cm in diameter). Eight artificial lip simulation devices are used to simulate eight puffs of a smoker taking a sweet and sour flavored cigarette. The tip of the sweet and sour flavored cigarette is inserted into the first artificial lip simulation device to a depth of 1 cm, dissolved for 2 seconds, then removed and left to stand for 30 seconds. It is then dissolved in the second artificial lip simulation device for 2 seconds, removed, and left to stand for 30 seconds. This process is repeated in the third artificial lip simulation device. Because the single-puff dissolution of characteristic components in the tip of the sweet and sour flavored cigarette is relatively low, in order to meet the requirements of instrumental quantitative analysis, five flavored cigarettes are taken for cumulative puff-by-puff dissolution according to the above operating procedure. After the dissolution test, the sponge was removed and added to a 50 mL centrifuge tube, along with 10 mL of internal standard aqueous solution. The mixture was then ultrasonically extracted at 20℃ and 40 kHz for 30 minutes to obtain eight dissolution samples for testing. The internal standard aqueous solution consisted of water, […]. 13Composed of C6-citric acid and sucralose-d6, both at a concentration of 10 ng / mL.

[0033] Liquid chromatography-tandem mass spectrometry detection: The sample of the dissolution solution to be tested was quantitatively detected by HPLC-MS / MS according to the liquid chromatography detection conditions and mass spectrometry detection conditions provided in Example 1. The results are shown in Table 5.

[0034] Table 5. Dissolution rate (mg / g) of characteristic components in sweet and sour flavored cigarette tipping paper 1 per puff Oral sequence Citric acid malic acid Sodium saccharin Sucralose Acesulfame K First bite 5.4600 3.2100 0.0600 2.6100 0.3600 Second mouth 1.2000 0.4815 0.0114 0.4437 0.0468 Third 0.8200 0.4710 0.0120 0.4300 0.0420 4th mouth 0.7300 0.4000 0.0100 0.4100 0.0400 5th 0.4200 0.3500 0.0090 0.3900 0.0380 6th 0.3500 0.3300 0.0000 0.3700 0.0360 7th 0.2100 0.2900 0.0000 0.3500 0.0300 8th 0.2000 0.2100 0.0000 0.3000 0.0280 Study on dissolution pattern: Table 5 shows that the characteristic components of sweet and sour flavored tipping paper 1 dissolve in a large amount in the first puff, and the dissolution of characteristic components decreases rapidly from the second puff onwards; this pattern is consistent with the actual sensory experience of smokers.

[0035] Therefore, the method provided in this embodiment of the invention enables the detection of puff-by-puff dissolution of characteristic components on sweet and sour flavored cigarette tipping paper and the study of puff-by-puff dissolution patterns. The sweet and sour characteristic components include acidulants: citric acid and malic acid; and sweeteners: acesulfame potassium, sodium saccharin, and sucralose. This embodiment of the invention uses artificial lip biomimicry to simulate the puff-by-puff dissolution of sweet and sour characteristic components during smoking, and simultaneously employs liquid chromatography-tandem mass spectrometry to achieve simultaneous detection of these components, improving detection efficiency and enabling the study of puff-by-puff dissolution patterns of characteristic components on sweet and sour flavored cigarette tipping paper. This provides technical support for quality monitoring during the production process of flavored cigarette tipping paper.

[0036] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them; although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications can still be made to the specific implementation of the present invention or equivalent substitutions can be made to some technical features without departing from the spirit of the technical solutions of the present invention, and all such modifications and substitutions should be covered within the scope of the technical solutions claimed in the present invention.

Claims

1. A method for detecting the puff-by-puff dissolution of characteristic components of sweet and sour flavored cigarette tipping paper, comprising the following steps: Sample pretreatment: Simulating the puff-by-puff dissolution of sweet and sour flavored cigarettes, multiple puff-by-puff dissolution samples were obtained for testing: among which, The characteristic components of the tipping paper for the sweet and sour flavored cigarettes include acidulants and sweeteners. The acidulants include citric acid and malic acid, and the sweeteners include acesulfame potassium, sodium saccharin, and sucralose. HPLC-MS / MS detection: The acidulant and sweetener in each of the test slurry samples were simultaneously and quantitatively detected using the HPLC-MS / MS method, wherein the HPLC-MS / MS method included the following liquid chromatography detection conditions: Liquid chromatography column: InfinityLab Poroshell 120 SB C 18 ; The mobile phase consists of an aqueous solution of ammonium acetate and methanol in a volume ratio of (0.8–10): (0–9.2), wherein the ammonium acetate in the aqueous solution is an additive. Elution method: gradient elution.

2. The detection method according to claim 1, characterized in that, The sample pretreatment steps include: using an artificial lip simulation device to simulate a smoker taking puffs of multiple sweet and sour flavored cigarettes, accumulating the dissolution corresponding to the number of puffs taken when the multiple sweet and sour flavored cigarettes are taken puff by puff, and obtaining multiple samples of the puff-by-puff dissolution to be tested.

3. The detection method according to claim 2, characterized in that, The sample pretreatment steps include: S1. Provides multiple artificial lip simulation devices, and each artificial lip simulation device includes a container and a sponge filled in the container and soaked in artificial saliva, wherein the sponge has a cigarette insertion hole in the middle that matches the diameter of the sweet and sour flavored cigarette. S2. Insert the tip of a sweet and sour flavored cigarette into the cigarette insertion holes of multiple artificial lip simulation devices in sequence, so that the characteristic components of the tip of the sweet and sour flavored cigarette dissolve into the corresponding sponge, and take out the sweet and sour flavored cigarette from the corresponding sponge to complete the dissolution experiment for a single sweet and sour flavored cigarette corresponding to the number of puffs, so that multiple sponges corresponding to the number of puffs contain the tip of the corresponding number of puffs dissolving liquid; S3. Repeat step S2 using multiple sweet and sour flavored cigarettes, and accumulate the corresponding number of sponge paper leachate in the sponge with the corresponding number of puffs. S4. Using water as the extraction solvent, extract the sponge containing the sponge with the corresponding number of suction ports to obtain multiple samples of the sponge to be tested.

4. The detection method according to claim 3, characterized in that, In step S4, an internal standard composition is added to the extraction solvent, and the internal standard composition includes... 13 C6-citric acid and sucralose-d6, in the sample of the dissolution solution to be tested 13 The concentrations of C6-citric acid and sucralose-d6 were both 5–10 ng / mL.

5. The detection method according to any one of claims 1-3, characterized in that, The column temperature of the liquid chromatography column is 25–45 °C, the concentration of the ammonium acetate aqueous solution is 2–10 mmol / L, and the gradient elution conditions are as follows: 。 6. The detection method according to claim 5, characterized in that, The column temperature is 40°C, and the gradient elution conditions are as follows: 。 7. The detection method according to any one of claims 4, characterized in that, The detection conditions also include: Flow rate: 0.1–0.5 mL / min; Injection volume: 8–12 μL.

8. The detection method according to claim 4, characterized in that, The HPLC-MS / MS method includes the following mass spectrometry detection conditions: electrospray ionization source, negative ion scanning mode; detection mode: multiple reaction monitoring.

9. The detection method according to claim 8, characterized in that, The mass spectrometry detection conditions also include: electrospray voltage: 4500 V; auxiliary gas pressure: 40 psi; ion source temperature: 400 ℃. The mass spectrometry parameters of the analyte are shown below: 。 10. A method for studying the puff-by-puff dissolution pattern of characteristic components in sweet and sour flavored cigarette tipping paper, comprising the following steps: Sample pretreatment: Simulating the puff-by-puff dissolution of sweet and sour flavored cigarettes, multiple puff-by-puff dissolution samples were obtained for testing: among which, The characteristic components of the tipping paper for the sweet and sour flavored cigarettes include acidulants and sweeteners. The acidulants include citric acid and malic acid, and the sweeteners include acesulfame potassium, sodium saccharin, and sucralose. HPLC-MS / MS detection: The acidulant and sweetener in each of the sip-by-sip dissolution samples were simultaneously quantitatively detected using the HPLC-MS / MS method to obtain the sip-by-sip dissolution amount of the acidulant and sweetener; wherein the HPLC-MS / MS method includes the following liquid chromatography detection conditions: Liquid chromatography column: InfinityLab Poroshell 120 SB C 18 ; The mobile phase consists of an aqueous solution of ammonium acetate and methanol in a volume ratio of (0.8–10): (0–9.2), wherein the ammonium acetate in the aqueous solution is an additive. Elution method: gradient elution; Regularity study: The dissolution rate of the acidulants and sweeteners was studied and analyzed to obtain the dissolution rate regularity of characteristic components of sweet and sour flavored cigarette tipping paper.

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