Method for measuring content of bisphenol A in tobacco substitute

Abstract

The invention provides a method for determining the content of bisphenol A in a tobacco substitute, and relates to the field of tobacco substitute detection. The method for measuring the content of bisphenol A in the tobacco substitute comprises the following steps: placing the tobacco substitute in an adsorbent, adding water for washing, and then extracting with an alcohol solvent to obtain a sample to be measured; the method comprises the following steps: introducing a sample to be detected into a high performance liquid chromatograph, separating through a chromatographic column, and detecting under an ultraviolet detector to obtain the content of bisphenol A in the tobacco substitute. The method provided by the invention solves the problems of lack of a standard method for testing the content of bisphenol A in the tobacco substitute, high detection cost, complex pretreatment, long time consumption and insufficient detection sensitivity, fills the blank of lack of a standard method for testing the content of bisphenol A in the tobacco substitute industry, and is beneficial to guarantee of consumer health. And the healthy development of tobacco substitutes is promoted.

Description

Technical Field

[0001] The present application relates to the field of tobacco substitute detection, and in particular to a method for determining the bisphenol A content in tobacco substitutes. Background Art

[0002] As an important chemical raw material, bisphenol A (BPA) is widely used in the synthesis of polycarbonate (PC), epoxy resin and other materials, which are further used to manufacture various daily necessities, such as plastic bottles, sippy cups for children, inner coatings of food packaging, and electronic cigarette accessories. The global production of plastics containing bisphenol A is as high as 27 million tons per year. The ubiquitous presence of bisphenol A in the environment and its potential harm to the human body have attracted widespread attention. Studies have shown that bisphenol A can interfere with the human endocrine system, cause damage to development and reproductive health, and is associated with a variety of serious health problems, such as breast cancer, prostate cancer, metabolic disorders and diabetes. Even very low doses of bisphenol A (10 -7 mol / L) can also cause the death of supporting cells and sperm cells, showing the serious harm of bisphenol A to the human body.

[0003] In the field of tobacco substitutes, due to the rapid development of the industry, the detection technology for various chemical substances in tobacco substitutes is still in the exploratory stage. Plastics in tobacco substitutes often contain materials containing bisphenol A such as PC. During the use of these materials, bisphenol A may migrate into tobacco substitutes and then be inhaled by consumers. Therefore, the testing of bisphenol A in tobacco substitutes is particularly important.

[0004] However, there is currently no standard method for testing the bisphenol A content in tobacco substitutes. The GC / MS method for determining bisphenol A requires derivatization treatment, and the pre-treatment steps are cumbersome and not conducive to rapid detection. Although the LC-MS / MS method has the advantages of strong qualitative ability, strong stability, and high sensitivity, its high detection cost limits its widespread application in ordinary laboratories. In addition, the detection method of high-performance liquid chromatography combined with fluorescence detector has the advantage of high sensitivity in the detection of bisphenol A, but its high cost and technical requirements constitute an obstacle in practical application. Summary of the invention

[0005] The purpose of this application is to provide a method for determining the bisphenol A content in tobacco substitutes to solve the above-mentioned problem.

[0006] A method for determining the content of bisphenol A in a tobacco substitute, comprising:

[0007] The tobacco substitute is placed in an adsorbent, washed with water, and then extracted with an alcohol solvent to obtain a sample to be tested;

[0008] Introduce the sample to be tested into a high-performance liquid chromatograph. After separation by a chromatographic column, detect it under an ultraviolet detector to obtain the content of bisphenol A in the tobacco substitute.

[0009] In some embodiments, the adsorbent includes a reversed-phase C18 solid-phase extraction cartridge;

[0010] And / or, the content of bisphenol A in the tobacco substitute is obtained by the following method: Use an ultraviolet detector to obtain the characteristic peak area of the sample to be tested, and substitute the characteristic peak area of the sample to be tested into the regression equation regarding the concentration of bisphenol A and the characteristic peak area to obtain the content of bisphenol A in the tobacco substitute.

[0011] In some embodiments, the regression equation regarding the concentration of bisphenol A and the characteristic peak area is obtained by the following method: Use a high-performance liquid chromatograph to detect multiple groups of bisphenol A standard products with different concentrations, and establish a standard curve and a regression equation regarding the concentration of bisphenol A and the characteristic peak area.

[0012] In some embodiments, the alcohol solvent includes methanol.

[0013] In some embodiments, the mass-volume ratio of the tobacco substitute to the water is 0.15 g / ml to 0.35 g / ml.

[0014] In some embodiments, the mass-volume ratio of the tobacco substitute to the alcohol solvent is 0.15 g / ml to 0.35 g / ml.

[0015] In some embodiments, after extraction with an alcohol solvent, the method further includes: successively subjecting the obtained extract to nitrogen blowing concentration, volume fixing, and filtration to obtain the sample to be tested.

[0016] In some embodiments, the detection conditions of the high-performance liquid chromatography include:

[0017] The chromatographic column includes a C18 chromatographic column;

[0018] The inner diameter of the chromatographic column is 4.6 mm, the column length of the chromatographic column is 250 mm, and the particle size of the chromatographic column is 4 μm;

[0019] The column temperature is 30 °C;

[0020] The flow rate is 1.0 ml / min;

[0021] The injection volume is 5 - 10 μL.

[0022] In some embodiments, the detection conditions of the high-performance liquid chromatography include:

[0023] The mobile phase includes mobile phase A and mobile phase B. Mobile phase A includes water, and mobile phase B includes methanol;

[0024] From 0 to 8 min, the volume fraction of mobile phase A changes from 45% to 30%, and the volume fraction of mobile phase B changes from 55% to 70%.

[0025] From 8 to 12 min, the volume fraction of mobile phase A changes from 30% to 20%, and the volume fraction of mobile phase B changes from 70% to 80%.

[0026] From 12 to 12.1 min, the volume fraction of mobile phase A changes from 20% to 10%, and the volume fraction of mobile phase B changes from 80% to 90%.

[0027] From 12.1 to 14 min, the volume fraction of mobile phase A is maintained at 10%, and the volume fraction of mobile phase B is maintained at 90%.

[0028] From 14 to 14.5 min, the volume fraction of mobile phase A changes from 10% to 45%, and the volume fraction of mobile phase B changes from 90% to 55%.

[0029] From 14.5 to 20 min, the volume fraction of mobile phase A is maintained at 45%, and the volume fraction of mobile phase B is maintained at 55%.

[0030] In some embodiments, the ultraviolet detector includes a diode array detector, and the detection wavelength of the diode array detector is 280 nm.

[0031] Compared with the prior art, the beneficial effects of the present application include:

[0032] The present application provides a method for determining the content of bisphenol A in tobacco substitutes with high efficiency, economy and high accuracy. The method of the present application solves the problems of lack of standard methods for testing the content of bisphenol A in tobacco substitutes, high detection costs, complex and time-consuming pretreatment, and insufficient detection sensitivity. The present application fills the blank of the lack of standard methods for testing the content of bisphenol A in the tobacco substitute industry, which is beneficial to protecting the health of consumers and promoting the healthy development of tobacco substitutes. Specifically, the method of the present application can efficiently extract bisphenol A from tobacco substitutes, effectively remove the interfering components in tobacco substitutes, and improve the accuracy of the method of the present application. Moreover, the present application uses high performance liquid chromatography ultraviolet method to determine the content of bisphenol A. Compared with GCMS method and liquid chromatography mass spectrometry method, the pretreatment operation of the method of the present application is simpler, significantly reducing the detection cost and improving the detection efficiency. In addition, the method of the present application also has the advantages of low detection limit and high sensitivity. In addition, the ultraviolet detector used in the method of the present application has the advantages of low price and wide applicability, which is beneficial to improving the applicability and popularity of the method of the present application. Description of the Drawings

[0033] To more clearly illustrate the technical solutions of the embodiments of the present application, the accompanying drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present application and should not be regarded as limiting the scope of the present application.

[0034] Figure 1 It is a flow chart of the method for determining the content of bisphenol A in tobacco substitutes in the present application;

[0035] Figure 2 It is the standard curve graph of bisphenol A constructed in Example 1 of the present application;

[0036] Figure 3 It is the chromatogram of the bisphenol A quality control sample;

[0037] Figure 4 It is the spectrogram of the bisphenol A quality control sample;

[0038] Figure 5 It is the chromatogram of the Tobacco-flavored spiked sample in Example 6. Specific embodiments

[0039] As used herein, the terms:

[0040] The terms "comprising", "including", "having", "containing" or any other variation thereof used herein are intended to cover non-exclusive inclusion. For example, a composition, step, method, article or device containing the listed elements is not necessarily limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article or device.

[0041] When an equivalent, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper preferred values and lower preferred values, this should be understood to specifically disclose all ranges formed by any pairing of any range upper limit or preferred value with any range lower limit or preferred value, regardless of whether the range is separately disclosed. For example, when the range "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5", etc. When a numerical range is described herein, unless otherwise stated, the range is intended to include its end values and all integers and fractions within the range.

[0042] The present application provides a method for determining the content of bisphenol A in tobacco substitutes, referring to Figure 1 , including:

[0043] Place the tobacco substitute in an adsorbent, rinse with water, and then extract with an alcohol solvent to obtain a sample to be tested;

[0044] The sample to be tested is introduced into a high performance liquid chromatograph. After separation by a chromatographic column, it is detected under an ultraviolet detector to obtain the content of bisphenol A in the tobacco substitute.

[0045] The components in the tobacco substitute are more complex than the matrices of many test samples, and interference is more likely to occur during the testing process. Bisphenol A is soluble in alcohol solvents and insoluble in water. The method of this application can efficiently extract bisphenol A from the tobacco substitute, and can remove many water-soluble interferences in the tobacco substitute. Compared with traditional ultrasonic extraction and shaking extraction, the method of this application can make the test matrix (sample to be tested) cleaner, have a higher detection limit, and less interference during the testing process.

[0046] In some embodiments, the adsorbent includes a reversed-phase C18 solid-phase extraction cartridge;

[0047] Further, the packing material of the C18 solid-phase extraction column includes Agilent SampliQ C18.

[0048] Specifically, in this application, the tobacco substitute is placed in a reversed-phase C18 solid-phase extraction cartridge, and the solid-phase extraction technology can be used to separate and enrich specific components in the tobacco oil. Further, flushing with water can wash away the water-soluble substances in the tobacco substitute. Since bisphenol A is insoluble in water, the step of flushing with water will not affect the content of bisphenol A. Bisphenol A can be dissolved in alcohol solvents. When extracted with alcohol solvents, bisphenol A will be eluted from the solid-phase extraction cartridge by the alcohol solvents, and other substances will remain on the solid-phase extraction column. In this process, the purpose of extracting bisphenol A from the tobacco substitute with complex components into the alcohol solvent can be achieved, realizing the separation of bisphenol A from other components in the tobacco substitute, and then the content of bisphenol A can be quantitatively detected by a high performance liquid chromatograph. The method of this application can accurately determine the content of bisphenol A in the tobacco substitute, providing important data support and relevant parameter verification data for the selection of each component and the addition of each substance in the tobacco substitute.

[0049] The minimum detectable solution concentration of the determination method of this application can reach 0.10 μg / mL, and it can accurately quantify trace amounts of bisphenol A in the tobacco substitute. Compared with the traditional HPLC ultraviolet method, the method of this application has the advantages of higher sensitivity and lower detection limit.

[0050] This application uses a liquid chromatograph to test the content of bisphenol A, and can monitor whether there is interference in the sample through the spectrogram. The probability of false positives is very low, greatly improving the qualitative accuracy of the determination of bisphenol A and having better selectivity.

[0051] In some embodiments, the content of bisphenol A in the tobacco substitute is obtained by the following method: using an ultraviolet detector to obtain the characteristic peak area of the sample to be measured, and substituting the characteristic peak area of the sample to be measured into the regression equation of bisphenol A concentration and characteristic peak area to obtain the content of bisphenol A in the tobacco substitute.

[0052] In some embodiments, the regression equation of bisphenol A concentration and characteristic peak area is obtained by the following method: using a high performance liquid chromatograph to detect multiple groups of bisphenol A standard products with different concentrations, and establishing a standard curve and a regression equation of bisphenol A concentration and characteristic peak area.

[0053] In some embodiments, the alcohol solvent includes methanol. Bisphenol A is easily soluble in methanol. Using methanol as an extraction reagent has a high extraction efficiency. Moreover, methanol is a common organic solvent with a low price, which is beneficial to reducing the cost of the method of the present application.

[0054] In some embodiments, the mass-volume ratio of the tobacco substitute to the water is 0.15 g / ml to 0.35 g / ml. When the mass-volume ratio of the tobacco substitute to the water is within the above range, the water can fully wet the filler in the solid phase extraction column, which is beneficial to effectively washing away the impurities soluble in water in the tobacco substitute.

[0055] For example, the mass-volume ratio of the tobacco substitute to the water can be 0.15 g / ml, 0.2 g / ml, 0.25 g / ml, 0.3 g / ml, 0.35 g / ml or any value between 0.15 g / ml and 0.35 g / ml.

[0056] In some embodiments, the mass-volume ratio of the tobacco substitute to the alcohol solvent is 0.15 g / ml to 0.35 g / ml. When the mass-volume ratio of the tobacco substitute to the alcohol solvent is within the above content range, the alcohol solvent can fully dissolve bisphenol A in the tobacco substitute to achieve efficient extraction.

[0057] For example, the mass-volume ratio of the tobacco substitute to the alcohol solvent can be 0.15 g / ml, 0.2 g / ml, 0.25 g / ml, 0.3 g / ml, 0.35 g / ml or any value between 0.15 g / ml and 0.35 g / ml.

[0058] In some embodiments, after extraction with an alcohol solvent, the method further includes: subjecting the obtained extract to nitrogen blowing for concentration, volume fixation, and filtration in sequence to obtain a sample to be measured. Since the extract contains a low concentration of bisphenol A, directly performing high performance liquid chromatography (HPLC) measurement may result in insufficient sensitivity or increased measurement error due to the low concentration. By nitrogen blowing for concentration, the concentration of bisphenol A in the extract can be increased, making it more suitable for the measurement range of HPLC and improving the accuracy and sensitivity of the measurement. Volume fixation is to adjust the concentrated extract to a fixed volume to ensure that each sample has the same volume during measurement, thereby eliminating the influence of volume differences on the measurement results. Filtration can remove possible fine particles or other insoluble impurities in the extract. The filtration operation can ensure that the solution entering the high performance liquid chromatograph is impurity-free, thereby improving the accuracy and reliability of the measurement. Exemplarily, filtration includes: filtering the volume-fixed sample solution to be measured through a 0.22 μm organic filter membrane.

[0059] In some embodiments, the detection conditions of the high performance liquid chromatography include:

[0060] The chromatographic column includes a chromatographic column; further, the C18 chromatographic column includes but is not limited to Poroshell 120SB-C18;

[0061] The inner diameter of the chromatographic column is 4.6 mm, the column length of the chromatographic column is 250 mm, and the particle size of the chromatographic column is 4 μm;

[0062] The column temperature is 30 °C;

[0063] The flow rate is 1.0 ml / min;

[0064] The injection volume is 5 - 10 μL. For example, the injection volume can be 5 μL, 6 μL, 7 μL, 8 μL, 9 μL, 10 μL or any value between 5 - 10 μL.

[0065] In some embodiments, the detection conditions of the high performance liquid chromatography include:

[0066] The mobile phase includes mobile phase A and mobile phase B. Mobile phase A includes water, and mobile phase B includes methanol;

[0067] From 0 to 8 min, the volume fraction of mobile phase A changes from 45% to 30%, and the volume fraction of mobile phase B changes from 55% to 70%;

[0068] From 8 to 12 min, the volume fraction of mobile phase A changes from 30% to 20%, and the volume fraction of mobile phase B changes from 70% to 80%;

[0069] 12 to 12.1 min, the volume fraction of mobile phase A changes from 20% to 10%, and the volume fraction of mobile phase B changes from 80% to 90%;

[0070] 12.1 to 14 min, the volume fraction of mobile phase A is maintained at 10%, and the volume fraction of mobile phase B is maintained at 90%; during this period, the high-concentration organic phase can flush the chromatographic column clean;

[0071] 14 to 14.5 min, the volume fraction of mobile phase A changes from 10% to 45%, and the volume fraction of mobile phase B changes from 90% to 55%;

[0072] 14.5 to 20 min, the volume fraction of mobile phase A is maintained at 45%, and the volume fraction of mobile phase B is maintained at 55%. This period is used to balance the pressure of the chromatographic column to ensure that it is fully prepared for the injection of the next sample.

[0073] In some embodiments, the ultraviolet detector includes a diode array detector, and the detection wavelength of the diode array detector is 280 nm.

[0074] The following will describe the implementation scheme of the present application in detail with specific embodiments. However, those skilled in the art will understand that the following embodiments are only used to illustrate the present application and should not be regarded as limiting the scope of the present application. For those not specified in the embodiments, they are carried out according to conventional conditions or conditions recommended by the manufacturer. For reagents or instruments not specified for the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0075] Example 1

[0076] Establish a standard curve:

[0077] Prepare bisphenol A standard solutions with concentrations of 0 μg / mL, 0.10 μg / mL, 0.20 μg / mL, 0.50 μg / mL, 1.0 μg / mL, 2.0 μg / mL, and 5.0 μg / mL. Use a high-performance liquid chromatograph to detect multiple groups of bisphenol A standard solutions with different concentrations, and establish a standard curve and regression equation for the concentration of bisphenol A and the characteristic peak area. The standard curve uses the concentration of bisphenol A as the abscissa and the characteristic peak area as the ordinate.

[0078] The parameter conditions of the high-performance liquid chromatograph are shown in Table 1, and the characteristic peak areas of bisphenol A at different concentrations are shown in Table 2.

[0079] Table 1 Parameter conditions of the high-performance liquid chromatograph

[0080]

[0081]

[0082] Table 2 Characteristic peak area table of bisphenol A at different concentrations

[0083]

[0084] According to the data in Table 2, combined with Figure 2 a standard curve of bisphenol A can be obtained, and the linear equation is Y = 4467.125X + 99.10412, where R 2 = 0.9999. It can be seen that the linearity of the bisphenol A standard curve is good, and R 2 is 0.9999, meeting the requirement of > 0.999.

[0085] Example 2

[0086] Example 2 provides a method for determining the content of bisphenol A in a tobacco substitute, including:

[0087] After shaking the tobacco substitute (e - liquid) evenly, weigh about 5.0 g of the tobacco substitute into a reversed - phase C18 solid - phase extraction cartridge; add 20 ml of ultrapure water and rinse repeatedly to remove water - soluble substances, and then extract the solid - phase extraction column with 20 ml of methanol;

[0088] Concentrate the extracted methanol by nitrogen blowing and make the volume constant to 1 ml, then pass it through a 0.22 - μm organic filter membrane into an injection vial to obtain a sample to be tested. Introduce the sample to be tested into a high - performance liquid chromatograph. After separation by the chromatographic column, detect it under an ultraviolet detector to obtain the characteristic peak area of the sample to be tested. Substitute the characteristic peak area of the sample to be tested into the regression equation of bisphenol A concentration and characteristic peak area to obtain the content of bisphenol A in the tobacco substitute.

[0089] Quality control sample detection: The purpose is to monitor and evaluate the accuracy of the experimental analysis method. The quality control sample contains e - liquid white oil (composed of nicotine, propylene glycol, and glycerol) and a bisphenol A standard. Add a 1 μg / ml bisphenol A standard to the e - liquid white oil. The detection method of the quality control sample is the same as the method for determining the content of bisphenol A in the tobacco substitute. Figure 3 is the chromatogram of the bisphenol A quality control sample. According to Figure 3 the characteristic peak area and regression equation in the chromatogram, the test result of bisphenol A is 0.975 μg / ml, and the standard deviation from the true value is 1.27%, meeting the strict requirement of standard deviation < 5%. Figure 4 is the spectrogram of the bisphenol A quality control sample. It can be seen from Figure 4 that bisphenol A has absorption at 280 nm.

[0090] Example 3: Recovery rate verification

[0091] Select 3 negative tobacco substitute samples as representative samples to verify the recovery rate and precision of bisphenol A.

[0092] The bisphenol A content in 3 negative tobacco substitute samples was determined, and the test results are shown in Table 3.

[0093] Table 3 Test results of 3 negative tobacco substitute samples

[0094]

[0095] As can be seen from Table 3, the bisphenol A content in the 3 negative tobacco substitute samples is 0.00 mg / kg, which are negative samples and can be used to verify the recovery rate and precision of bisphenol A.

[0096] The above 3 - flavored negative tobacco substitute samples were spiked respectively from three aspects of low, medium, and high concentrations. After being processed according to the steps of Example 2 and then determined, the data in Table 4 below were obtained.

[0097] Table 4 Recovery rate table of bisphenol A at different spiked concentrations

[0098]

[0099] As can be seen from Table 4, the recovery rate range of bisphenol A is: 98% - 106%, meeting the requirements of 90% - 110%. This shows that in tobacco substitutes with such a complex matrix, the high - performance liquid chromatography - ultraviolet method can reduce the influence of the matrix on the determination of bisphenol A and greatly improve the accuracy.

[0100] Example 4: Precision verification

[0101] An equal amount of bisphenol A solution with the same concentration was added to 6 negative tobacco substitute samples respectively. After being processed according to the steps of Example 2 and then determined, the data in Table 5 were obtained.

[0102] Table 5 Relative standard deviation RSD (%) of bisphenol A at the same concentration

[0103]

[0104] As can be seen from Table 5, the RSD of bisphenol A is 0.98% - 3.74%, meeting the requirement of < 10%, indicating that the reproducibility of this method is good.

[0105] Example 5: Selectivity verification

[0106] Three - flavored negative tobacco substitute samples in Example 3 were selected, a certain amount of bisphenol A standard solution was added, and after being processed according to the steps in Example 2 and then determined, qualitative and quantitative analysis was carried out based on the retention time and peak area, and the interference situation of the sample was checked by combining with the absorption spectrogram to judge the selectivity.

[0107] The tobacco substitute with Strawberry Ice flavor was treated with this method and then tested by HPLC. The peak shape was good, and there was no interference peak within the retention time ±0.3min, so the bisphenol A content could be accurately measured.

[0108] The tobacco substitute with Strawberry Banana flavor was treated with this method and then tested by HPLC. The peak shape was good, and there was no interference peak within the retention time ±0.3min, so the bisphenol A content could be accurately measured.

[0109] The tobacco substitute with peach berry flavor was treated with this method and then tested by HPLC. The peak shape was good, and there was no interference peak within the retention time ±0.3min, so the bisphenol A content could be accurately measured.

[0110] Based on the above three flavors of tobacco substitute samples, it is preliminarily judged that the method of this application has good selectivity.

[0111] Example 6

[0112] A method for determining the content of bisphenol A in a tobacco substitute comprises the following steps:

[0113] S1: Randomly select an e-cigarette with a flavor of Tobacco, and the test result is 0.00mg / kg.

[0114] S2: Disassemble the above-mentioned flavored electronic cigarettes, take out the oil storage cotton, put it into a clean sample bag, and pour the e-liquid into a clean sampling vial;

[0115] S3: Weigh 5.0 g of the e-liquid sample of the flavor and put it into a reverse phase C18 solid phase extraction column; add 20 ml of ultrapure water to repeatedly rinse off the water-soluble substances, and then extract the solid phase extraction column with 20 ml of methanol;

[0116] S4: The extracted methanol is concentrated to 1 ml by nitrogen blowing and then filtered through a 0.22 μm organic filter membrane for HPLC testing;

[0117] Tobacco flavored e-liquid was treated with this method and then tested by HPLC. Figure 5 As shown, the peak shape is good, there is no interference peak within the retention time ±0.3min, and no interference is observed in the absorption spectrum, so the bisphenol A content can be accurately measured. Therefore, it can be concluded that the tobacco flavored e-liquid treated by this method and tested on the machine has strong anti-interference ability and good selectivity.

[0118] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the various embodiments of the present application.

[0119] In addition, those skilled in the art will appreciate that although some embodiments herein include certain features included in other embodiments but not others, the combination of features of different embodiments means that it is within the scope of the present application and forms different embodiments. For example, in the claims above, any one of the claimed embodiments can be used in any combination. The information disclosed in this background section is only intended to enhance the understanding of the overall background of the present application and should not be regarded as an admission or any form of suggestion that this information constitutes prior art already known to those skilled in the art.

Claims

1. A method for determining the content of bisphenol A in a tobacco substitute, characterized in that: include: The tobacco substitute is placed in an adsorbent, washed with water, and then extracted with an alcohol solvent to obtain a sample to be tested; The sample to be tested is introduced into a high performance liquid chromatograph, separated by a chromatographic column, and then detected under an ultraviolet detector to obtain the bisphenol A content in the tobacco substitute.

2. The method for determining the content of bisphenol A in tobacco substitutes according to claim 1, characterized in that: The adsorbent includes a reverse phase C18 solid phase extraction cartridge; And / or, the bisphenol A content in the tobacco substitute is obtained by: using an ultraviolet detector to obtain the characteristic peak area of ​​the sample to be tested, substituting the characteristic peak area of ​​the sample to be tested into a regression equation regarding bisphenol A concentration and characteristic peak area, and obtaining the bisphenol A content in the tobacco substitute.

3. The method for determining the content of bisphenol A in tobacco substitutes according to claim 2, characterized in that: The regression equation for bisphenol A concentration and characteristic peak area is obtained by the following method: using a high performance liquid chromatograph to detect multiple groups of bisphenol A standards of different concentrations, and establishing a standard curve and a regression equation for bisphenol A concentration and characteristic peak area.

4. The method for determining the content of bisphenol A in tobacco substitutes according to claim 1, characterized in that: The alcohol solvent includes methanol.

5. The method for determining the content of bisphenol A in tobacco substitutes according to claim 1, characterized in that: The mass volume ratio of the tobacco substitute to the water is 0.15 g / ml to 0.35 g / ml.

6. The method for determining the content of bisphenol A in a tobacco substitute according to claim 1, characterized in that: The mass volume ratio of the tobacco substitute to the alcohol solvent is 0.15 g / ml to 0.35 g / ml.

7. The method for determining the content of bisphenol A in a tobacco substitute according to claim 1, characterized in that: After extraction with an alcohol solvent, the method further comprises: nitrogen blowing, concentrating, volume fixing and filtering the obtained extract in sequence to obtain a sample to be tested.

8. The method for determining the content of bisphenol A in tobacco substitutes according to claim 1, characterized in that: The detection conditions of the high performance liquid chromatography include: The chromatographic column includes a C18 chromatographic column; The inner diameter of the chromatographic column is 4.6 mm, the column length of the chromatographic column is 250 mm, and the particle size of the chromatographic column is 4 μm; The column temperature was 30°C; Flow rate: 1.0 ml / min; The injection volume is 5-10 μL.

9. The method for determining the content of bisphenol A in a tobacco substitute according to claim 8, characterized in that: The detection conditions of the high performance liquid chromatography include: The mobile phase includes a mobile phase A and a mobile phase B, wherein the mobile phase A includes water and the mobile phase B includes methanol; From 0 to 8 min, the volume fraction of the mobile phase A changes from 45% to 30%, and the volume fraction of the mobile phase B changes from 55% to 70%; 8-12 min, the volume fraction of the mobile phase A changes from 30% to 20%, and the volume fraction of the mobile phase B changes from 70% to 80%; 12-12.1 min, the volume fraction of the mobile phase A changes from 20% to 10%, and the volume fraction of the mobile phase B changes from 80% to 90%; 12.1-14 min, the volume fraction of the mobile phase A is maintained at 10%, and the volume fraction of the mobile phase B is maintained at 90%; 14-14.5 min, the volume fraction of the mobile phase A changes from 10% to 45%, and the volume fraction of the mobile phase B changes from 90% to 55%; From 14.5 to 20 min, the volume fraction of the mobile phase A is maintained at 45%, and the volume fraction of the mobile phase B is maintained at 55%.

10. The method for determining the content of bisphenol A in a tobacco substitute according to claim 1, characterized in that: The ultraviolet detector comprises a diode array detector, and the detection wavelength of the diode array detector is 280nm.

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