A method for simultaneously detecting caffeine, menthone, quassine and reineckite A in electronic cigarette atomization

Abstract

The application provides a method for simultaneously detecting caffeine, menthone, quassine and reineckite A in electronic cigarette atomization, and belongs to the technical field of electronic cigarette detection. The method comprises the following steps: step one, using 50% methanol solution as a solvent to extract the electronic cigarette atomization to obtain a solution to be detected; caffeine, menthone, quassine and reineckite A standard substances are respectively taken to prepare standard working solutions; step two, using a liquid chromatograph to perform liquid phase analysis and detection on the solution to be detected and the standard working solutions to obtain the content of caffeine, menthone, quassine and reineckite A in the electronic cigarette atomization. The method provided by the application can simultaneously detect caffeine, menthone, quassine and reineckite A in the electronic cigarette atomization, and has the advantages of high result accuracy and repeatability and strong durability.

Description

Technical Field

[0001] This invention relates to the field of electronic cigarette detection and analysis technology, and in particular to a method for simultaneously detecting caffeine, menthone, quassin, and calciferoside A in electronic cigarette atomized products. Background Technology

[0002] E-cigarettes, as non-combustible electronic products that mimic cigarettes, have similar effects to regular cigarettes, providing alertness, satisfying nicotine cravings, and producing feelings of euphoria and relaxation. They are also tar-free, making them widely popular among smokers. E-liquid, also known as e-cigarette liquid, is the electronic atomizing liquid used with e-cigarettes. Heated by an e-cigarette atomizer, it produces a vapor similar to that of cigarettes. The main components of e-liquid are food-grade or pharmaceutical-grade glycerin, 1,2-propanediol, polyethylene glycol, and tobacco-specific flavorings, and it also contains nicotine. Substandard e-liquids can produce harmful chemicals when inhaled, such as caffeine, menthol, quassin, and calcein A.

[0003] Caffeine is a neuroactive alkaloid widely found in tea, beverages, and coffee. Recently, public awareness of e-cigarette safety has increased, and many countries and organizations have clear regulations on the amount of caffeine added to e-cigarette liquids. For example, the EU Tobacco Products Directive (TPD), the French Afnor e-cigarette standard, and the American E-cigarette Industry Association (AEMSA) group standard T / CECC 002-2021 explicitly prohibit the addition of caffeine to e-cigarette liquids. my country's e-cigarette group standard also lists caffeine as one of the prohibited components in e-cigarettes.

[0004] Mentholone is an extract component of peppermint plants. Oral administration may cause liver cancer, lung tissue deformation, and growths in mice. Mentholone is classified as a Group 2B carcinogen by the World Health Organization, and the U.S. Food and Drug Administration banned its use as a food additive last year. There is a potential risk of excessive levels of this ingredient in peppermint-flavored and menthol-flavored e-cigarette products.

[0005] Quercusin, also known as bitter quinolone, is a bioactive triterpenoid extracted from the stem bark of a plant; stachysin A, also known as stachysin A or stachysin glycoside A, is a plant extract. Menthone, quercusin, and stachysin A are listed together as prohibited or restricted substances in Appendix B of EN 17648:2022, "Ingredients of E-cigarette Liquids".

[0006] Currently, the standard for caffeine detection in the e-cigarette industry is mostly GB 5009.139—2014, "Determination of Caffeine in Beverages." This method often requires the addition of reagents such as magnesium oxide and trichloroacetic acid in the pretreatment process to eliminate interference from proteins and pigments. It is suitable for complex matrices such as beverages and coffee / tea products, but the pretreatment involves relatively complex steps such as purification and impurity removal. Furthermore, this method can only be used for the single-item detection of caffeine. Existing technologies also provide other methods for detecting caffeine, such as Chinese invention patent application number 201711277083.9, which is an automated detection method for caffeine in e-cigarette liquids. The disadvantages of this method are that it only tests for a single item, and the testing process requires liquid-liquid extraction, making it difficult to guarantee accuracy and recovery rate. Another example is Chinese invention patent application number 202111266984.4, which is a method for detecting vanillin and caffeine in e-cigarette liquids. This method involves ultrasonic extraction of e-cigarette samples to obtain extracted samples, followed by gas chromatography analysis of the samples to be tested. This method has simple pretreatment, but the test items are only vanillin and caffeine. At the same time, the simple gas chromatography method is prone to false positives and cannot perform accurate qualitative analysis.

[0007] There are currently no reference detection methods for three substances in e-cigarette e-liquids, flavorings, additives, etc.: menthone, quassin, and calciferoside A. Furthermore, no existing technology has been found that can simultaneously detect these four substances in e-cigarette e-liquids. Summary of the Invention

[0008] To address the aforementioned issues, this invention aims to provide a rapid, efficient, simple, and accurate detection method with high recovery rate, enabling the simultaneous detection of caffeine, menthone, quassin, and calciferoside A in electronic cigarette smoke compounds.

[0009] On the one hand, this application provides a method for simultaneously detecting caffeine, menthone, quassin, and calciferoside A in electronic cigarette vapors, comprising:

[0010] Step 1: Extract the e-cigarette atomized substances using a 50% methanol aqueous solution as the solvent to obtain the solution to be tested; prepare standard working solutions by taking caffeine, menthol, quassin, and calciferoside A standard substances respectively.

[0011] Step 2: Perform liquid chromatography analysis on the test solution and the standard working solution to obtain the contents of caffeine, menthol, quassin, and calciferoside A in the electronic cigarette smoke.

[0012] Among them, the UV detection channel corresponding to cadaverine A is 220nm, the UV detection channel corresponding to quassin and menthone is 254nm, and the UV detection channel corresponding to caffeine is 272nm.

[0013] In the method provided in this application, the wavelength of the detection channel corresponds to the absorption wavelength of the characteristic peak generated by ultraviolet spectral absorption in liquid chromatography analysis. The wavelength setting of the detection channel can be achieved by adjusting the wavelength setting of the ultraviolet detector in the liquid chromatograph.

[0014] In this application, the electronic atomizing compound refers to the raw material inhaled in electronic cigarette products.

[0015] In one embodiment, the ratio of electronic smoke atomizer and 50% methanol aqueous solution in step one is (0.1-1)g:(10-15)mL.

[0016] In one embodiment, the extraction step in step one includes: vortex oscillation treatment for 1 to 3 minutes, ultrasonic treatment for 15 to 30 minutes, and filtration membrane.

[0017] In one embodiment, the rotational speed of the vortex oscillation treatment is 2000 rpm; and / or, the frequency of the ultrasonic treatment is 40-60 kHz; and / or, the pore size of the filter membrane is 0.22 μm.

[0018] In one embodiment, the standard working solution in step one is prepared using anhydrous methanol as a solvent; and / or, the concentration range of the standard working solution is 0.5–50 mg / L.

[0019] In one embodiment, in the liquid phase analysis detection step of step two, the mobile phase is a water-methanol solution, and gradient elution is performed using a 20%–70% water-30%–80% methanol solution by volume percentage.

[0020] In one embodiment, the elution procedure of the gradient elution step is as follows: 70% water + 30% methanol, 0-3 min; 50% water + 50% methanol, 3-8 min, hold for 6 min; 20% water + 80% methanol, 14-16 min, hold for 4 min; 70% water + 30% methanol, 20-20.1 min, hold for up to 25 min.

[0021] In one embodiment, the liquid phase analysis detection step in step two uses a PDA detector, a C18 column, an injection volume of 10 μL, a column temperature of 30 °C, and a flow rate of 1.0 mL / min.

[0022] In one embodiment, the test water is Grade I water, and the methanol purity is required to be chromatographically pure or higher.

[0023] It is understood that the specific operational steps of the liquid chromatography analysis in the detection method provided in this application can adopt the operational steps of existing liquid chromatography instruments, including experimental operation procedures, computer software operation procedures, and calculation methods for the content of the components to be detected in the sample.

[0024] In one embodiment, the content X of caffeine, menthone, quassin, and calciferoside A in the sample, expressed in mg / kg, is calculated using the following formula:

[0025]

[0026] In the formula:

[0027] C i The concentrations of caffeine, menthol, quassin, and calciferoside A in the sample are indicated (unit: mg / L); C o V represents the concentration of caffeine, menthol, quassin, and calciferoside A in the blank (unit: mg / L); V represents the total volume of the sample (unit: mL); f represents the dilution factor of the sample; and m represents the sample mass (unit: g).

[0028] The test results are expressed as the arithmetic mean of parallel measurements, rounded to two decimal places.

[0029] In one embodiment, the detection limits for caffeine, menthone, quassin, and calciferoside A in the detection method provided in this application are:

[0030] Caffeine (CAS No.: 58-08-2), detection limit: 1.0 mg / kg;

[0031] Mentholone (CAS No.: 89-82-7), detection limit: 1.0 mg / kg;

[0032] Quercusin (CAS No.: 76-78-8), detection limit: 2.0 mg / kg;

[0033] Calcinin A (CAS No.: 12798-51-5), Detection limit: 2.0 mg / kg.

[0034] On the other hand, this application also provides the application of the method for simultaneously detecting caffeine, menthol, quassin, and calciferoside A in electronic cigarette atomizers in the quality testing of electronic cigarette products.

[0035] In one embodiment, the e-cigarette atomizer comprises e-cigarette liquid, tobacco flavoring, and / or tobacco additives.

[0036] This application has at least the following beneficial effects:

[0037] 1. This invention provides a liquid chromatography method for the simultaneous detection of caffeine, menthol, quassin, and calcitonin A in e-cigarette atomized products. This method has the advantages of high accuracy, repeatability, and robustness. Furthermore, in the detection method of this application, the sample to be tested is extracted with a 50% methanol aqueous solution, which can simultaneously extract caffeine, menthol, quassin, and calcitonin A from flavorings and e-cigarette atomized products. The pretreatment steps are simple and the recovery rate is high.

[0038] 2. This invention employs high-performance liquid chromatography-ultraviolet spectroscopy for detection. Detection wavelengths of 220 nm, 254 nm, and 272 nm are selected based on the characteristic absorption wavelengths of caffeine, menthone, quassin, and cadaverine A, respectively. A specific gradient elution program is used for chromatographic separation, resulting in good resolution and anti-interference capabilities. Especially for quassin and cadaverine A, which are difficult to separate, excellent separation results are achieved, improving the feasibility and accuracy of the detection method.

[0039] 3. This invention eliminates the need for testing personnel to come into contact with highly toxic organic reagents, ensuring human health and safety. It is also environmentally friendly, simple to operate, and suitable for rapid and efficient quality testing in the industrial production of e-cigarette products. Attached Figure Description

[0040] The accompanying drawings, which are incorporated in and form part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0041] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort. In the drawings:

[0042] Figure 1 The full-wavelength scan chromatograms of the four mixed standards in Example 1 obtained by instrument method 1;

[0043] Figure 2 The full-wavelength scan chromatograms obtained by testing the four mixed standards in Example 1 using instrumental method 2 are shown below.

[0044] Figure 3 The ultraviolet spectrum of the caffeine standard substance in Example 2;

[0045] Figure 4 The ultraviolet spectrum of the menthol standard substance in Example 2;

[0046] Figure 5 The ultraviolet spectrum of the quassin standard substance in Example 2;

[0047] Figure 6 The ultraviolet spectrum of the cadaverine A standard substance in Example 2;

[0048] Figure 7 The image shows the ultraviolet spectrum of the mixed standard obtained at the extraction wavelength of 220 nm in Example 2.

[0049] Figure 8 The ultraviolet spectrum of the mixed standard obtained at the extraction wavelength of 254 nm in Example 2 is shown.

[0050] Figure 9 The ultraviolet spectrum of the mixed standard obtained at the extraction wavelength of 272 nm in Example 2 is shown.

[0051] Figure 10 The above are liquid chromatograms of the four standard substances in Example 2: caffeine, menthol, quassin, and calciferoside A, corresponding to the 220nm, 254nm, and 272nm detection channels from top to bottom, respectively.

[0052] Figure 11 This is an extraction rate curve of four substances under different extraction solvents in Example 4. Detailed Implementation

[0053] To more clearly illustrate the overall concept of this application, a detailed description is provided below by way of embodiments. Numerous specific details are set forth in the following description to provide a more thorough understanding of this application. However, it will be apparent to those skilled in the art that this application can be implemented without one or more of these details. In other instances, to avoid confusion with this application, some technical features well-known in the art have not been described.

[0054] Unless otherwise specified, in the following embodiments, reagents or instruments whose manufacturers are not indicated are all conventional products that can be purchased commercially.

[0055] Unless otherwise specified in the examples, the conditions shall be performed according to the standard conditions or the conditions recommended by the manufacturer.

[0056] Example 1

[0057] This embodiment optimizes the gradient elution procedure for the detection of caffeine, menthol, quassin, and calciferoside A. The specific steps are as follows:

[0058] Weigh out a certain amount of caffeine, menthol, quassin, and calciferoside A standard substances, dissolve them completely in methanol, and then test them using a liquid chromatograph.

[0059] Instrumentation and Method 1: Liquid Chromatography: Waters e2695 with PDA detector; Column: Waters SunFire C18, 5μm, 4.6*250mm; Injection volume: 10μl; Column temperature: 30℃; Flow rate: 1.0mL / min; Mobile phase: A-water, B-methanol; Gradient elution program: 70%A + 30%B (0-3min) → 20%A + 80%B (3-10min, hold for 5min) → 70%A + 30%B (15-15.1min, hold for 20min).

[0060] Instrument Method 2: Liquid Chromatography: Waters e2695 with PDA detector; Column: Waters SunFire C18, 5μm, 4.6*250mm; Injection volume: 10μl; Column temperature: 30℃; Flow rate: 1.0mL / min; Mobile phase: A-water, B-methanol; Gradient elution program: 70%A+30%B (0-3min) → 50%A+50%B (3-8min, hold for 6min) → 20%A+80%B (14-16min, hold for 4min) → 70%A+30%B (20-20.1min, hold for 25min).

[0061] The four mixed standard substances—caffeine, menthol, quassin, and calciferoside A—were tested separately using the instrumental method described above, yielding full-wavelength scanning chromatograms. Figure 1 , Figure 2 .like Figure 1 As shown, the separation of quassin and calciferoside A in the chromatogram obtained by instrumental method 1 was not satisfactory. Therefore, based on instrumental method 1, the elution program was optimized to obtain instrumental method 2. Compared with the gradient elution program of instrumental method 1, instrumental method 2 adds a 50% water + 50% methanol gradient elution, which is more conducive to the separation of the four compounds, such as... Figure 2 As shown, quassin and calciferoside A have good separation effects in the chromatogram obtained by instrument method 2.

[0062] Example 2

[0063] This embodiment optimizes the detection channels selected for the detection of caffeine, menthol, quassin, and calciferoside A. The specific steps are as follows:

[0064] Weigh out a certain amount of caffeine, menthol, quassin, and calciferoside A standard substances, dissolve them completely in methanol, and then test them using a liquid chromatograph.

[0065] The four mixed standard substances—caffeine, menthol, quassin, and calciferoside A—were tested according to instrument method 2 in Example 1. The obtained spectral information is as follows: Figure 3-6 As shown. According to Figure 3-6It can be seen that the four substances have strong characteristic absorption peaks near 272nm (caffeine), 254nm (quassin and menthone), and 220nm (calciferoside A), respectively.

[0066] The chromatograms of the four mixed standard substances were extracted at wavelengths of 220 nm, 254 nm, and 272 nm to obtain the chromatograms. Figure 7-9 ,according to Figure 7-9 As shown, at a detection wavelength of 220 nm, cadaverine A exhibits a strong absorption peak response, while quassin and menthol show weaker responses; at a detection wavelength of 254 nm, quassin and menthol show strong responses, but cadaverine A shows a weaker response; at a detection wavelength of 272 nm, caffeine shows a strong response, but quassin, menthol, and cadaverine A show relatively low responses. Considering all factors, this method selects three different absorption wavelengths as detection channels: 272 nm (caffeine), 254 nm (quassin and menthol), and 220 nm (cadaverine A).

[0067] Four mixed standard substances were detected using these three detection wavelengths to obtain the detection chromatogram. Figure 10 .like Figure 10 As shown, through detection channel optimization, this method exhibits good separation performance for the four substances, effectively eliminating interference from related substances, especially quassinolide and calciferoside A, two substances with similar retention rates. Specificity refers to the ability of a method to accurately determine the characteristics of the analyte in the presence of other components, reflecting the method's accurate and specific determination capability for the test analyte in the presence of coexisting substances. Therefore, this method demonstrates good specificity.

[0068] Example 3

[0069] This embodiment verifies the linearity of the detection method, and the specific steps are as follows:

[0070] Standard curve: Weigh out specific amounts of caffeine, menthol, quassin, and calcein A standards into brown volumetric flasks, dissolve completely in methanol, and dilute to volume to prepare primary standard stock solutions. Transfer five aliquots of each primary standard stock solution to prepare working standard solutions with concentrations of 0.5 mg / L, 1 mg / L, 2 mg / L, 5 mg / L, and 10 mg / L, respectively.

[0071] The treated samples were tested according to instrument method 2 in Example 1. The linearity of caffeine, menthol, quassin, and calciferoside A at various concentrations is shown in Table 1:

[0072] Table 1

[0073]

[0074] As shown in Table 1, the linear correlation coefficients of the four substances obtained by this test method are all R0. 2 A value ≥0.999 indicates that the detection method provided in this application has a good linear relationship.

[0075] Example 4

[0076] This embodiment optimizes the sample processing method, and the specific steps are as follows:

[0077] Sample preparation: 50g of e-cigarette oil (provided by Shenzhen Zhenwei Biotechnology Co., Ltd.) was selected. Since the e-cigarette oil does not contain caffeine, menthol, quassin, and calcitonin A, certain amounts of caffeine, menthol, quassin, and calcitonin A standard substances were added to prepare a positive sample with theoretical contents of 26.0mg / kg caffeine, 90.0mg / kg menthol, 18.0mg / kg quassin, and 13mg / kg calcitonin A: e-cigarette vaporized product (1#).

[0078] Pretreatment: Weigh 0.5g of each of the same electronic cigarette smoke compound (1#) into 15mL centrifuge tubes, then add pure water, 100% methanol, 100% ethanol, 100% acetonitrile, 30% methanol-water, 50% methanol-water, and 70% methanol-water to a final volume of 10mL. Vortex the tubes for 2min, then sonicate them for 20min in an ultrasonic extractor with a frequency range of 50kHz. After extraction, filter the samples through a 0.22μm filter membrane to obtain 8 samples to be tested.

[0079] Instrumental testing: The processed samples were tested according to instrumental method 2 in Example 1. The content X of caffeine, menthol, quassin, and calciferoside A in the sample, expressed in mg / kg, was calculated using the following formula:

[0080]

[0081] In the formula:

[0082] C i The concentrations of caffeine, menthol, quassin, and calciferoside A in the sample are indicated (unit: mg / L); C o V represents the concentration of caffeine, menthol, quassin, and calciferoside A in the blank (unit: mg / L); V represents the total volume of the sample (unit: mL); f represents the dilution factor of the sample; and m represents the sample mass (unit: g).

[0083] The test results are expressed as the arithmetic mean of parallel determinations, rounded to two decimal places. Relevant test results and extraction rate results are shown in Table 2 and... Figure 11 As shown:

[0084] Table 2 Sample Test Values

[0085]

[0086]

[0087] Combine Table 2 and Figure 11 The results show that methanol extraction is slightly better than ethanol and acetonitrile extraction, and mixed extraction solvents are significantly better than pure solvent extraction. The optimal extraction solvent ratio is 50:50 for methanol and water.

[0088] Example 5

[0089] This embodiment verifies the precision of the detection method, and the specific steps are as follows:

[0090] Pretreatment: Weigh 0.5g of each of the six portions of electronic smoke compound (1#) from Example 4 into 15mL centrifuge tubes, then add 50% methanol water to make up to 10mL, vortex on a vortex shaker for 2min, and then place in an ultrasonic extractor for 20min. The ultrasonic extractor frequency range is 50kHz. After extraction, the samples are filtered through a 0.22μm filter membrane to obtain six samples to be tested.

[0091] Instrument testing: The processed samples were tested according to instrument method 2 in Example 1. The relevant test results are shown in Table 3 below:

[0092] Table 3 Sample Test Values

[0093]

[0094]

[0095] As shown in Table 3, when tested according to this method, the precision is 1.3% (n=6) for caffeine, 0.4% (n=6) for menthone, 0.8% (n=6) for quassin, and 1.0% (n=6) for cadaverine A. Precision refers to the degree of similarity between results obtained from multiple measurements of the same homogeneous sample under specified test conditions. Therefore, this test method exhibits good precision when simultaneously testing caffeine, menthone, quassin, and cadaverine A.

[0096] Example 6

[0097] This embodiment verifies the accuracy of the detection method, and the specific steps are as follows:

[0098] Pretreatment: Weigh 12 portions of the same electronic cigarette smoke compound (2#) and place each 0.5g into a 15mL centrifuge tube. Perform a three-concentration, three-parallel spiking experiment on 9 of the samples, with spiking amounts of 5μg, 20μg, and 100μg, respectively. Then add 50% methanol water to make up to 10mL, vortex on a vortex shaker for 2min, and then place in an ultrasonic extractor for 20min. The frequency range of the ultrasonic extractor is 50kHz. After extraction, filter the sample through a 0.22μm filter membrane to obtain 12 samples to be tested.

[0099] Instrument testing: The processed samples were tested according to instrument method 2 in Example 1, and the relevant test results are shown in Table 4.

[0100] Table 4 Sample Test Values

[0101]

[0102] Spike recovery rate = (sample spiked test value × sample weight - sample test value × sample weight) / spiked amount × 100%. The spike recovery results are shown in Table 5.

[0103] Table 5 Spike Recovery Results

[0104]

[0105] Combining the results in Tables 4 and 5, the recoveries of different concentrations of each item when tested according to this method are as follows: caffeine 80.2%–104.9%, menthol 83.2%–105.2%, quassin 89.8%–101.6%, and calciferoside A 91.6%–105.2%. Accuracy refers to the degree of agreement between the test results and the receiving reference value. This experiment used the spiked recovery rate to verify the accuracy of the method. The above results show that this test method has good accuracy when simultaneously testing caffeine, menthol, quassin, and calciferoside A.

[0106] Example 7

[0107] Pretreatment: Weigh 0.5g of each of the same electronic cigarette smoke compound (1#) into 15mL centrifuge tubes, add 50% methanol water to make up to 10mL, vortex on a vortex shaker for 2min, and then place in an ultrasonic extractor for 20min. The ultrasonic extractor frequency range is 50KHZ. After extraction, filter the sample through a 0.22μm filter membrane. Repeat this operation 6 times to obtain 6 samples to be tested.

[0108] Instrument Method 3: Chromatographic column: ZORBAX Eclipse Plus C18, specifications: 5μm 4.6*250mm, injection volume: 10μl; column temperature: 30℃; flow rate: 1.0mL / min; mobile phase: A-water, B-methanol; gradient elution program: 70%A+30%B (0-3min) → 50%A+50%B (3-8min) → 50%A+50%B (8-14min) → 20%A+80%B (14-16min) → 20%A+80%B (16-20min) → 70%A+30%B (20-20.1min, hold for 25min).

[0109] Instrumental testing: The processed samples were tested according to the instrumental method 3 described above, and the test results were compared with those of Example 5. The relevant comparison results are shown in Table 6 below:

[0110] Table 6 Comparison Results

[0111]

[0112] The data comparison in Table 6 shows that when testing according to this method, even when using different brands of chromatographic columns, the test results still have good precision, and the test results of different brands of chromatographic columns show good consistency. This indicates that this test method has good robustness when simultaneously testing for caffeine, menthol, quassin, and calcein A.

[0113] The above description is merely an embodiment of this application and is not intended to limit the scope of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of the claims of this application.

Claims

1. A method for simultaneously detecting caffeine, menthol, quassin, and calciferoside A in e-cigarette vapors, characterized in that, include: Step 1: Extract the e-cigarette atomized substances using a 50% methanol aqueous solution as a solvent to obtain the solution to be tested; Standard working solutions were prepared by taking caffeine, menthol, quassin, and calciferoside A standard substances respectively; Step 2: Perform liquid chromatography analysis on the test solution and the standard working solution to obtain the contents of caffeine, menthol, quassin, and calciferoside A in the electronic cigarette smoke. In the liquid chromatography detection step, a C18 column was used, and gradient elution was performed using a 20%~70% water-30%~80% methanol solution (by volume percentage). The elution program was as follows: 70% water + 30% methanol, 0~3 min; 50% water + 50% methanol, 3~8 min, hold for 6 min; 20% water + 80% methanol, 14~16 min, hold for 4 min; 70% water + 30% methanol, 20~20.1 min, hold for up to 25 min. Among them, the UV detection channel corresponding to cadaverine A is 220nm, the UV detection channel corresponding to quassin and menthone is 254nm, and the UV detection channel corresponding to caffeine is 272nm.

2. The method according to claim 1, characterized in that, In step one, the ratio of electronic smoke atomizer to 50% methanol aqueous solution is (0.1~1) g : (10~15) mL.

3. The method according to claim 1, characterized in that, The extraction steps in step one include: vortex oscillation treatment for 1-3 minutes, ultrasonic treatment for 15-30 minutes, and filtration membrane.

4. The method according to claim 3, characterized in that, The rotational speed of the vortex oscillation treatment is 2000 rpm; and / or the frequency of the ultrasonic treatment is 40~60 kHz; and / or the pore size of the filter membrane is 0.22 μm.

5. The method according to claim 1, characterized in that, In step one, the standard working solution is prepared using anhydrous methanol as the solvent; and / or, the concentration range of the standard working solution is 0.5~50 mg / L.

6. The method according to claim 1, characterized in that, In the liquid phase analysis detection step of step two, a PDA detector is used, the injection volume is 10 μL, the column temperature is 30℃, and the flow rate is 1.0 mL / min.

7. The application of the method for simultaneously detecting caffeine, menthone, quassin, and calciferoside A in electronic cigarette atomized products as described in any one of claims 1-6 in the quality testing of electronic cigarette products.

8. The application according to claim 7, characterized in that, The e-cigarette atomizers include e-cigarette liquids and / or tobacco additives.

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