CN108760917B - Tobacco and tobacco product treatment method and method for measuring content of N-nitrosodiethanolamine in tobacco and tobacco product - Google Patents
Tobacco and tobacco product treatment method and method for measuring content of N-nitrosodiethanolamine in tobacco and tobacco product Download PDFInfo
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Abstract
The invention relates to a processing method of tobacco and tobacco products and a measuring method of N-nitrosodiethanolamine content in the tobacco and the tobacco products, belonging to the technical field of harmful component detection in the tobacco and the tobacco products. The invention relates to a method for processing tobacco and tobacco products, which comprises the following steps: extracting a to-be-detected product by adopting an alkaline salting-out agent solution, then adding an organic solvent for extraction, and layering; adding a silane derivatization reagent into the organic phase or adding the silane derivatization reagent into the organic phase after the organic phase volatilizes the solvent, and obtaining a sample solution after the N-nitrosodiethanolamine completely reacts. The treatment method of the invention transfers the extracted N-nitrosodiethanolamine from the water phase to the organic phase, and then the N-nitrosodiethanolamine reacts with the silane derivatization reagent, thereby avoiding the adsorption of hydroxyl polar groups in a chromatographic column and improving the sensitivity of the N-nitrosodiethanolamine in detection.
Description
Technical Field
The invention relates to a processing method of tobacco and tobacco products and a measuring method of N-nitrosodiethanolamine content in the tobacco and the tobacco products, belonging to the technical field of harmful component detection in the tobacco and the tobacco products.
Background
N-nitrosodiethanolamine is a non-volatile nitrosamine, which is toxic and classified as a class 2 human carcinogen by the International agency for research on cancer (IARC), and is also classified as a harmful substance list in tobacco and tobacco products in the guideline draft for harmful and potentially harmful substances in tobacco products and cigarette smoke issued by the FDA tobacco products center in 2012. Therefore, the method for determining the content of N-nitrosodiethanolamine in the tobacco and the tobacco products has important significance for risk assessment of the tobacco and the tobacco products. According to literature research, although the quantity of literature related to nitrosamine in tobacco is large, the literature mainly focuses on the detection of tobacco-specific nitrosamine and non-polar volatile nitrosamine, and the existing literature is less concerned with the detection of N-nitrosodiethanolamine in tobacco. In the fields of cosmetics, environment and the like, a relatively large number of documents are reported in view of the toxicity and carcinogenicity of N-nitrosodiethanolamine, but liquid chromatography-based methods such as HPLC-UV and HPLC-MS/MS are mainly adopted, and solid phase extraction is also required for purification. For example, Wangfeng et al HPLC-MS/MS measures NDELA in cosmetics, and the quantitative limit of this method is 50 ng/g. There are also a few documents which adopt GC-TEA (gas chromatography-thermal energy analysis combined instrument) to detect N-nitrosodiethanolamine in matrixes such as cosmetics, but TEA is not possessed by conventional laboratories, has low instrument popularity, also needs to introduce a solid phase extraction step, and cannot eliminate the adsorption problem of N-nitrosodiethanolamine on chromatographic columns and liner tubes. The N-nitrosodiethanolamine has low response on mass spectrum, and is easy to adsorb on a chromatographic column to cause peak shape broadening due to the existence of hydroxyl polar groups, so that the detection sensitivity is lower than two orders of magnitude of volatile nitrosamine and tobacco-specific nitrosamine, and the current situation that the GC-MS/MS cannot be directly adopted to directly detect the N-nitrosodiethanolamine is caused.
Disclosure of Invention
The invention aims to provide a method for treating tobacco and tobacco products, which can improve the sensitivity of N-nitrosodiethanolamine in detection.
The invention also provides a method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products.
In order to achieve the above object, the technical scheme adopted by the method for treating tobacco and tobacco products of the invention is as follows:
a method of treating tobacco and tobacco products comprising the steps of: extracting a to-be-detected product by adopting an alkaline salting-out agent solution, then adding an organic solvent for extraction, and layering; adding a silane derivatization reagent into the organic phase or adding the silane derivatization reagent into the organic phase after the organic phase volatilizes the solvent, and obtaining a sample solution after the N-nitrosodiethanolamine completely reacts.
The tobacco and the processing method of the tobacco product are suitable for processing tobacco shreds, tobacco leaves and smokeless tobacco products, wherein the smokeless tobacco product can be any one of bagged mouth tobacco, bulk mouth tobacco, chewing tobacco, buccal tobacco and gum base type tobacco products. In the method for treating the tobacco and the tobacco products, the N-nitrosodiethanolamine is extracted from the tobacco and the tobacco products by adopting an alkaline salting-out agent solution, the extracted N-nitrosodiethanolamine is transferred from a water phase to an organic phase by adopting an organic solvent which is not mutually soluble with water, and then the N-nitrosodiethanolamine is reacted with a silane derivatization reagent, so that the adsorption of a hydroxyl polar group in a chromatographic column is avoided, and the sensitivity of the N-nitrosodiethanolamine in detection is improved; particularly, the organic solvent in the organic phase is volatilized, the volatile components in the organic phase can be removed, the interference of impurity components on the detection result is reduced, and the detection sensitivity is further improved.
The basic salting-out agent solution is a basic salting-out agent aqueous solution. Preferably, the sample is extracted with an alkaline salting-out agent solution by immersing the sample in the alkaline salting-out agent solution. The soaking time is 5-20 min.
The alkaline salting-out agent solution is alkaline saturated salting-out agent solution.
OH in the basic salting-out agent solution-The concentration of the ions is 0.2 to 1.5 mol/L.
The salting-out agent in the alkaline salting-out agent solution is sodium chloride, potassium chloride, ammonium sulfate, sodium sulfate and potassium sulfate.
Preferably, when the sample is extracted by using the basic salting-out agent solution, the volume of the basic salting-out agent solution used per 0.5-1 g of the sample is 2-5 mL.
Preferably, the basic salting-out agent solution is a saturated salting-out agent solution containing 1-5 wt% of sodium hydroxide.
The organic solvent is immiscible with water. Preferably, the organic solvent is at least one of diethyl ether, dichloromethane, methyl tert-butyl ether and chloroform.
The volume ratio of the alkaline salting-out agent solution to the organic solvent is 2-5: 5-10.
The extraction is vortex oscillation. The vortex oscillation time is 20-40 min.
The method for treating the tobacco and the tobacco products further comprises the step of centrifuging after adding the organic solvent for extraction. Solid-liquid separation can be accelerated and realized through centrifugation, the layering of an organic phase and a water phase is promoted, and the extraction residues dispersed in the organic phase after layering are reduced. The centrifugal rotating speed is 3000-10000 r/min.
The temperature when the organic solvent is volatilized is 20-35 ℃. Because the temperature of the volatile solvent is not high, in order to accelerate the speed of the volatile solvent, a blow-drying mode can be adopted.
Preferably, the method for treating tobacco and tobacco products further comprises removing water from the organic phase before adding the silane derivatization reagent into the organic phase or before volatilizing the solvent from the organic phase. Water removal can be carried out by adding an inorganic solid desiccant to the organic phase. Adding an inorganic solid drying agent into the organic phase, oscillating, standing, and then taking the organic phase as a silane derivatization reagent or taking the organic phase as a volatile solvent and then adding the silane derivatization reagent.
The inorganic solid desiccant is a neutral inorganic solid desiccant. The inorganic solid desiccant is at least one of anhydrous sodium sulfate, anhydrous calcium chloride and anhydrous magnesium sulfate.
The silane derivatizing agent is added in an amount sufficient to completely react the N-nitrosodiethanolamine in the organic phase taken. Conventional silane derivatizing agents may be used in the present invention.
Preferably, the silane derivatizing agent is at least one of N, O-bis (trimethylsilyl) trifluoroacetamide, N-methyl-N- (trimethylsilyl) trifluoroacetamide.
Preferably, the reaction temperature is 35-60 ℃. The reaction time is 30-60 min.
Extracting the to-be-detected product by adopting an alkaline salting-out agent solution to obtain an extracting solution and extracting slag mixed with the extracting solution. The extraction residue is retained in the extraction solution, and when the extraction solution is uniformly mixed with the organic solvent, the N-nitrosodiethanolamine in the water phase can be transferred into the organic phase, so that the concentration of the N-nitrosodiethanolamine in the water phase is reduced, and the dissolution of the N-nitrosodiethanolamine in the water phase is promoted.
The invention relates to a method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products,
a method for detecting N-nitrosodiethanolamine in tobacco and tobacco products comprises the following steps:
1) extracting the to-be-detected product added with the internal standard substance by adopting an alkaline salting-out agent solution, then adding an organic solvent for extraction, and layering; adding a silane derivatization reagent into the organic phase or adding the silane derivatization reagent into the organic phase after the organic phase volatilizes the solvent, and obtaining a sample solution after the N-nitrosodiethanolamine completely reacts;
2) the content of N-nitrosodiethanolamine in the sample solution is determined by gas chromatography-mass spectrometry.
The method for measuring the content of the N-nitrosodiethanolamine in the tobacco and the tobacco products can effectively detect the N-nitrosodiethanolamine in the tobacco and the tobacco products, has good chromatographic peak shape, high sensitivity, good reproducibility, small matrix interference and high recovery rate, can meet the detection requirement of a gas chromatography-mass spectrometry method of the N-nitrosodiethanolamine in the tobacco and the tobacco products, is suitable for measuring a large batch of samples, and can provide reference for the detection of the N-nitrosodiethanolamine in other sample systems such as food, plants, environment and the like.
The internal standard substance is N-nitrosodiethanolamine-d 8. The internal standard substance is added into the to-be-detected product, namely, the internal standard substance is firstly dissolved in water to prepare an internal standard solution, and then the internal standard solution is added into the to-be-detected product. The concentration of the internal standard solution is 50-500 ng/mL. The volume of the internal standard solution correspondingly adopted by each 0.5-1 g of the to-be-detected product is 20-100 mu L.
The basic salting-out agent solution is a basic salting-out agent aqueous solution. Preferably, the sample is extracted with an alkaline salting-out agent solution by immersing the sample in the alkaline salting-out agent solution. The soaking time is 5-20 min.
The alkaline salting-out agent solution is alkaline saturated salting-out agent solution.
OH in the basic salting-out agent solution-The concentration of the ions is 0.2 to 1.5 mol/L.
The salting-out agent in the alkaline salting-out agent solution is sodium chloride, potassium chloride, ammonium sulfate, sodium sulfate and potassium sulfate.
Preferably, when the sample is extracted by using the basic salting-out agent solution, the volume of the basic salting-out agent solution used per 0.5-1 g of the sample is 2-5 mL.
Preferably, the basic salting-out agent solution is a saturated salting-out agent solution containing 1-5 wt% of sodium hydroxide.
The organic solvent is immiscible with water. Preferably, the organic solvent is at least one of diethyl ether, dichloromethane, methyl tert-butyl ether and chloroform.
The volume ratio of the alkaline salting-out agent solution to the organic solvent is 2-5: 5-10.
The extraction is vortex oscillation. The vortex oscillation time is 20-40 min.
The method for treating the tobacco and the tobacco products further comprises the step of centrifuging after adding the organic solvent for extraction. Solid-liquid separation can be accelerated by centrifugation, and extraction residues dispersed in the organic phase after layering are reduced. The centrifugal rotating speed is 3000-10000 r/min.
The temperature when the organic solvent is volatilized is 20-35 ℃. Because the temperature of the volatile solvent is not high, in order to accelerate the speed of the volatile solvent, a blow-drying mode can be adopted.
Preferably, the method for treating tobacco and tobacco products further comprises removing water from the organic phase before adding the silane derivatization reagent into the organic phase or before volatilizing the solvent from the organic phase. Water removal can be carried out by adding an inorganic solid desiccant to the organic phase. Adding an inorganic solid drying agent into the organic phase, oscillating, standing, and then taking the organic phase as a silane derivatization reagent or taking the organic phase as a volatile solvent and then adding the silane derivatization reagent.
The inorganic solid desiccant is a neutral inorganic solid desiccant. The inorganic solid desiccant is at least one of anhydrous sodium sulfate, anhydrous calcium chloride and anhydrous magnesium chloride.
The silane derivatizing agent is added in an amount sufficient to completely react the N-nitrosodiethanolamine in the organic phase taken. Conventional silane derivatizing agents may be used in the present invention.
Preferably, the silane derivatizing agent is at least one of N, O-bis (trimethylsilyl) trifluoroacetamide (i.e., BSTFA), N-methyl-N- (trimethylsilyl) trifluoroacetamide (i.e., MSTFA).
Preferably, the reaction temperature is 35-60 ℃. The reaction time is 30-60 min.
The gas chromatography-mass spectrometry adopts GC-MS/MS for detection.
The chromatographic conditions for GC-MS/MS detection are as follows: flow rate of carrier gas: 1.0-2.0 mL/min; sample inlet temperature: 200 to 280 ℃; sample introduction amount: 1 mu L of the solution; the split ratio is 2: 1-10: 1; temperature rising procedure: keeping the temperature at 50 ℃ for 2min, then increasing the temperature to 280 ℃ at a heating rate of 10 ℃/min, and keeping the temperature for 10 min.
The chromatographic column adopted by a gas chromatograph in GC-MS/MS detection comprises the following steps: DB-5 (length 30 m-60 m, inner diameter 0.25mm, film thickness 0.25 μm).
Carrier gas adopted by a gas chromatograph in GC-MS/MS detection: and (e) He.
The mass spectrum conditions of GC-MS/MS detection are as follows: EI ionization source, MRM mode; the quantitative ion pair of N-nitrosodiethanolamine was 130/116, and the ionization energy was 15 eV.
When N-nitrosodiethanolamine-d 8 is used as an internal standard substance, the mass spectrum conditions further comprise: the quantitative ion of N-nitrosodiethanolamine-d 8 was 136/119, and the ionization energy was 15 eV.
Drawings
FIG. 1 is a flowchart of a method for measuring the content of N-nitrosodiethanolamine in example 1;
FIG. 2 is the MRM spectrum of N-nitrosodiethanolamine in bulk buccal tobacco samples and standards in example 1.
Detailed Description
The technical solution of the present invention will be further described with reference to the following embodiments.
Example 1
The method for measuring the content of N-nitrosodiethanolamine in the tobacco and the tobacco product of the embodiment, as shown in fig. 1, includes the following steps:
1) grinding bulk buccal tobacco under the condition of adding liquid nitrogen, weighing 0.5g of ground sample, transferring to a 50mL centrifuge tube, adding 50 μ L of internal standard solution (100ng/mL of N-nitrosodiethanolamine-d 8 solution), adding 5mL of saturated sodium chloride aqueous solution containing 2% by mass of sodium hydroxide, and standing for 15 min;
2) after standing, adding 10mL of ethyl acetate into a centrifugal tube, carrying out vortex oscillation for 20min, then centrifuging for 5min at 6000r/min, and layering; adding 2g of anhydrous sodium sulfate into 5mL of the upper organic phase, uniformly mixing, standing for 1h, putting 200 mu L of supernate into a 5mL centrifuge tube, adding 200 mu L of BSTFA, and reacting for 30min in a water bath at 60 ℃ to obtain a sample solution;
3) detecting N-nitrosodiethanolamine in the sample solution by GC-MS/MS, wherein the result is shown in figure 2; the GC-MS/MS parameters were as follows:
a chromatographic column: DB-5(30m 0.25mm 0.25 μm); carrier gas: he, flow rate: 2.0 ml/min; sample inlet temperature: 280 ℃; sample introduction amount: 1 mu L of the solution; the split ratio is 10: 1; temperature rising procedure: keeping the temperature at 50 ℃ for 2min, then increasing the temperature to 280 ℃ at the heating rate of 10 ℃/min, and keeping the temperature for 10 min;
MS/MS parameters: EI ionization source (ion source temperature is 300 ℃, quadrupole temperature is 180 ℃), and MRM mode detection is adopted, wherein the quantitative ion pair of N-nitrosodiethanolamine is 130/116, and the ionization energy is 15 ev; the quantitative ion of N-nitrosodiethanolamine-d 8 is 136/119, and the ionization energy is 15 ev;
4) respectively preparing N-nitrosodiethanolamine standard solutions with different concentrations, drawing a working curve by using the peak area to the concentration under the condition of GC-MS/MS detection in the step 3), and obtaining a detection limit and a quantification limit by using 5 times of signal-to-noise ratio as the quantification limit, wherein the working curve is shown in Table 1.
TABLE 1 working curves
| Linear equation of equations | Coefficient of linearity (R) | Linear range (ng/g) | Detection limit (ng/g) | Quantitative limit (ng/g) |
| Y＝5.234X-0.032 | 0.9991 | 0.5-50 | 0.1 | 0.3 |
Example 2
The method for measuring the content of N-nitrosodiethanolamine in the tobacco and the tobacco products comprises the following steps:
1) directly taking 1 bag of bagged buccal cigarette A (the mass is about 1g) as a to-be-measured product, and weighing; then tearing the bagged buccal cigarette A from the middle, transferring the buccal cigarette powder to a 50mL centrifuge tube, placing the torn bag in the 50mL centrifuge tube, adding 100 muL of internal standard solution (50ng/mL of N-nitrosodiethanolamine-d 8 solution), adding 5mL of saturated sodium chloride aqueous solution containing 5 percent (by weight) of sodium hydroxide, and standing for 20 min;
2) after standing, adding 12mL of ethyl acetate into a centrifugal tube, carrying out vortex oscillation for 15min, then centrifuging for 3min at 5000r/min, and layering; adding 3g of anhydrous sodium sulfate into 8mL of the upper organic phase, standing for 1h, putting 100 mu L of supernatant into a 5mL centrifuge tube, adding 200 mu L of BSTFA, and reacting for 30min in a water bath at 60 ℃ to obtain a sample solution;
3) detecting N-nitrosodiethanolamine in the sample solution by GC-MS/MS, and calculating the content of the N-nitrosodiethanolamine according to the working curve in the table 1; the GC-MS/MS parameters were the same as in example 1.
5 times of parallel experiments are carried out according to the method for measuring the content of the N-nitrosodiethanolamine in the tobacco and the tobacco products in the embodiment, the measured precision (N is 5) is 5.6 percent, and the reproducibility of the method is proved.
Example 3
The method for measuring the content of N-nitrosodiethanolamine in the tobacco and the tobacco products comprises the following steps:
1) directly weighing 1.0g of tobacco shred powder sample, transferring to a 50mL centrifuge tube, adding 50 μ L of internal standard solution (40ng/mL of N-nitrosodiethanolamine-d 8 solution), adding 5mL of saturated sodium chloride aqueous solution containing 5% sodium hydroxide, and standing for 20 min;
2) after standing, adding 12mL of dichloromethane into a centrifugal tube, carrying out vortex oscillation for 15min, then centrifuging for 3min at 5000r/min, and layering; taking 8mL of lower organic phase, adding 3g of anhydrous sodium sulfate, standing for 1h, taking 200 mu L of supernatant, placing the supernatant into a 5mL centrifuge tube, adding 200 mu L of MSTFA, and reacting for 30min in a water bath at 37 ℃ to obtain a sample solution;
3) detecting N-nitrosodiethanolamine in the sample solution by GC-MS/MS, and calculating the content of the N-nitrosodiethanolamine according to the working curve in the table 1; the GC-MS/MS parameters were as in example 1 and found to be 2.3 ng/g.
Example 4
The method for measuring the content of N-nitrosodiethanolamine in the tobacco and the tobacco products comprises the following steps:
1) grinding bulk buccal tobacco under the condition of adding liquid nitrogen, weighing 0.5g of ground sample, transferring to a 50mL centrifuge tube, adding 50 μ L of internal standard solution (100ng/mL of N-nitrosodiethanolamine-d 8 solution), adding 2mL of saturated sodium chloride aqueous solution containing 1% (by mass) of sodium hydroxide, and standing for 5 min;
2) after standing, adding 10mL of ethyl acetate into a centrifugal tube, carrying out vortex oscillation for 40min, centrifuging for 5min at 3000r/min, and layering; adding 2g of anhydrous sodium sulfate into 5mL of the upper organic phase, uniformly mixing, standing for 1h, putting 1.5mL of supernatant into a 5mL centrifuge tube, drying at normal temperature, adding 500 mu L of BSTFA, and reacting in a water bath at 45 ℃ for 45min to obtain a sample solution;
3) detecting N-nitrosodiethanolamine in the sample solution by GC-MS/MS, and calculating the content of the N-nitrosodiethanolamine according to the working curve in the table 1; the GC-MS/MS parameters were as in example 1 and found to be 1.8 ng/g.
Experimental example 5
This example is a spiking recovery test, the test method comprising the steps of:
1) placing 3 50mL centrifuge tubes in parallel, and respectively marking as a centrifuge tube A, a centrifuge tube B and a centrifuge tube C; weighing 3 bags of bagged buccal cigarettes B respectively, wherein the bagged buccal cigarettes B do not contain N-nitrosodiethanolamine;
2) 1 bag of bagged buccal cigarette B (the mass is about 1g) is respectively placed into each centrifuge tube, the bagged buccal cigarette B is torn from the middle when the buccal cigarette B is placed into each centrifuge tube, the buccal cigarette powder is transferred into the centrifuge tube, and the torn bags are also placed into the same centrifuge tube; then, respectively and sequentially adding 100 mu L of internal standard solution (50ng/mL of N-nitrosodiethanolamine-d 8 solution), N-nitrosodiethanolamine standard solution and 5mL of saturated sodium chloride aqueous solution containing 5% of sodium hydroxide into each centrifuge tube, and standing for 20 min; the amounts of the N-nitrosodiethanolamine standard sample added into the centrifugal tube A, B, C are respectively low, medium and high, the addition amount in the centrifugal tube A is 2ng/g, the addition amount in the centrifugal tube B is 4ng/g, and the addition amount in the centrifugal tube C is 8 ng/g;
3) after standing, adding 12mL of ethyl acetate into a centrifuge tube A, carrying out vortex oscillation for 15min, centrifuging for 3min at the speed of 5000r/min, and layering; adding 3g of anhydrous sodium sulfate into 8mL of the upper organic phase, standing for 1h, putting 100 mu L of supernatant into a 5mL centrifuge tube, adding 200 mu L of BSTFA, and reacting for 30min in a water bath at 60 ℃ to obtain a sample solution A;
performing the above treatment on the centrifugal tube B and the centrifugal tube C to respectively obtain a sample solution B and a sample solution C;
4) respectively sampling a sample solution A, a sample solution B and a sample solution C, and detecting N-nitrosodiethanolamine in the sample solution A, the sample solution B and the sample solution C by adopting GC-MS/MS; the GC-MS/MS parameters were the same as in example 1.
Based on the measurement results, the recovery rates of the spiked samples were calculated, respectively, and the results are shown in Table 2.
TABLE 2 measurement results of N-nitrosodiethanolamine in bagged buccal tobacco sample B and recovery rate with standard
| Compound (I) | Content (wt.) | Recovery rate (Low) | Recovery rate (middle) | Recovery rate (high) |
| N-nitrosodiethanolamine | 0ng/g | 81％ | 102％ | 106％ |
As can be seen from the data in Table 2, the determined recovery rate is between 81 and 106 percent, which shows that the analysis method has high accuracy.
Claims (6)
1. A method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products is characterized in that: the method comprises the following steps:
1) extracting the to-be-detected product added with the internal standard substance by adopting an alkaline salting-out agent solution, then adding an organic solvent for extraction, and layering; adding a silane derivatization reagent into the organic phase or adding the silane derivatization reagent into the organic phase after the organic phase volatilizes the solvent, and obtaining a sample solution after the N-nitrosodiethanolamine completely reacts;
OH in the basic salting-out agent solution-The concentration of the ions is 0.2-1.5 mol/L; the salting-out agent in the alkaline salting-out agent solution is sodium chloride, potassium chloride, ammonium sulfate, sodium sulfate and potassium sulfate; when the to-be-detected product is extracted by adopting an alkaline salting-out agent solution, the volume of the alkaline salting-out agent solution adopted per 0.5-1 g of the to-be-detected product is 2-5 mL; the volume ratio of the alkaline salting-out agent solution to the organic solvent is 2-5: 5~10；
2) Measuring the content of N-nitrosodiethanolamine in a sample solution by adopting a gas chromatography-mass spectrometry method;
the gas chromatography-mass spectrometry adopts GC-MS/MS for detection; the chromatographic conditions for GC-MS/MS detection are as follows: a chromatographic column: DB-5; flow rate of carrier gas: 1.0-2.0 mL/min; sample inlet temperature: 200-280 ℃; sample introduction amount: 1 mu L of the solution; the split ratio is 2: 1-10: 1; temperature rising procedure: keeping the temperature at 50 ℃ for 2min, then increasing the temperature to 280 ℃ at the heating rate of 10 ℃/min, and keeping the temperature for 10 min; the mass spectrum conditions of GC-MS/MS detection are as follows: EI ionization source, MRM mode.
2. The method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products according to claim 1, wherein: the method also comprises the step of carrying out water removal treatment on the organic phase before adding a silane derivatization reagent into the organic phase or before taking a volatile solvent from the organic phase.
3. The method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products according to claim 1, wherein: the silane derivatization reagent is at least one of N, O-bis (trimethylsilyl) trifluoroacetamide and N-methyl-N- (trimethylsilyl) trifluoroacetamide.
4. The method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products according to claim 1, wherein: the organic solvent is at least one of diethyl ether, dichloromethane, methyl tert-butyl ether and chloroform.
5. The method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products according to claim 1, wherein: the extraction is vortex oscillation.
6. The method for measuring the content of N-nitrosodiethanolamine in tobacco and tobacco products according to claim 1, wherein: the reaction temperature is 35-60 ℃.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101963604A (en) * | 2010-10-29 | 2011-02-02 | 川渝中烟工业公司 | Method for measuring sterol in tobaccos |
| CN104359999A (en) * | 2014-11-03 | 2015-02-18 | 上海烟草集团有限责任公司 | Method for measuring tobacco-specific N-nitrosamine in tobacco |
| CN105353068A (en) * | 2015-12-09 | 2016-02-24 | 中国烟草总公司郑州烟草研究院 | GC-MS/MS assay determination method for volatile N-nitrosamine in tobacco and smokeless tobacco products |
| CN105891383A (en) * | 2016-06-04 | 2016-08-24 | 中国烟草总公司郑州烟草研究院 | GC-MS/MS method for synchronously detecting specific nitrosamine and volatile N-nitrosamine in tobacco and tobacco products |
| CN106501410A (en) * | 2016-10-27 | 2017-03-15 | 安徽易康达光电科技有限公司 | The detection method of N nitrosodiethanolamines in a kind of cosmetics |
-
2018
- 2018-05-24 CN CN201810508897.7A patent/CN108760917B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101963604A (en) * | 2010-10-29 | 2011-02-02 | 川渝中烟工业公司 | Method for measuring sterol in tobaccos |
| CN104359999A (en) * | 2014-11-03 | 2015-02-18 | 上海烟草集团有限责任公司 | Method for measuring tobacco-specific N-nitrosamine in tobacco |
| CN105353068A (en) * | 2015-12-09 | 2016-02-24 | 中国烟草总公司郑州烟草研究院 | GC-MS/MS assay determination method for volatile N-nitrosamine in tobacco and smokeless tobacco products |
| CN105891383A (en) * | 2016-06-04 | 2016-08-24 | 中国烟草总公司郑州烟草研究院 | GC-MS/MS method for synchronously detecting specific nitrosamine and volatile N-nitrosamine in tobacco and tobacco products |
| CN106501410A (en) * | 2016-10-27 | 2017-03-15 | 安徽易康达光电科技有限公司 | The detection method of N nitrosodiethanolamines in a kind of cosmetics |
Non-Patent Citations (4)
| Title |
|---|
| Analysis of N-Nitrosodiethanolamine(NDELA) in Cosmetics, Shampoos, and Raw Materials by Gas Chromatography-Mass Fragmentography;Birendra n. et al.;《28th Annual Conference on Mass Spectrometry and Allied Topics》;19800530;第364-365页 * |
| DETERMINATION OF N-NITROSODIETHANOLAMINE IN COSMETICS HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY AND GAS CHROMATOGRAPHY-MASS SPECTROMETRY AS ALTERNATIVE METHODS TO CHEMILUMINESCENCE DETECTION;ROLF SCHWARZENBACH et al.;《Journal of Chromatography》;19891231;第231-242页 * |
| GC-TEA测定化妆品中的NDELA;邢昆明 等;《广州化工》;20161231;第84-88页 * |
| N-Nitrosodiethanolamine in tobacco: method validation and levels present in US tobacco products https://www.coresta.org/sites/default/files/abstracts/2012_TSRC54_Williamson.pdf;WILLIAMSON F. et al.;《Tob. Sci. Res. Conf.》;https://www.coresta.org/sites/default/files/abstracts/2012_TSRC54_Williamson.pdf;20121231;第1-27页 * |
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