Method for measuring TSNAs (tobacco-specific nitrosamines) in electronic cigarette smoke

A measurement method and technology of electronic cigarettes, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of few reports on measurement methods, and achieve the effects of simple pre-treatment, improved analysis throughput, and accurate analysis results

Active Publication Date: 2014-12-24
CHINA TOBACCO YUNNAN IND
10 Cites 12 Cited by

AI-Extracted Technical Summary

Problems solved by technology

Therefore, the detection of TSNAs content in electronic cigarette smoke is of great signific...
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Abstract

The invention relates to a high performance liquid chromatography-tandem mass spectrometry method for measuring TSNAs (tobacco-specific nitrosamines) in electronic cigarette smoke, which belongs to the technical field of physical and chemical inspection of an electronic cigarette. The method is characterized by comprising the steps: sucking an electronic cigarette sample by virtue of a smoking machine, trapping the electronic cigarette smoke by adopting a full-smoke trapping device, adopting a Cambridge filter to be serially connected with an absorption bottle, independently adopting the Cambridge filter or independently adopting the absorption bottle, measuring the smoke by adopting a high efficiency liquid phase chromatography-triple quadrupole tandem mass spectrograph, and quantifying analyte by adopting an internal standard method. The method is a brand new method for measuring the content of TSNAs in the electronic cigarette smoke. The method is good in selectivity and accuracy.

Application Domain

Component separation

Technology Topic

Tandem mass spectrometryNitrosamine +7

Image

  • Method for measuring TSNAs (tobacco-specific nitrosamines) in electronic cigarette smoke
  • Method for measuring TSNAs (tobacco-specific nitrosamines) in electronic cigarette smoke
  • Method for measuring TSNAs (tobacco-specific nitrosamines) in electronic cigarette smoke

Examples

  • Experimental program(1)

Example Embodiment

[0031] Example 1:
[0032] A liquid chromatography tandem mass spectrometry method for the determination of tobacco peculiar nitrosamines in electronic cigarette smoke. The test process is that the electronic cigarette smoke sample is diluted and directly introduced into a high performance liquid chromatography-triple quadrupole tandem mass spectrometry system for analysis.
[0033] This method is the determination of four specific nitrosamines: NNK, NNN, NAT and NAB, and includes the following steps:
[0034] a. Cigarette smoking: e-cigarette samples are smoked in Canada deep smoking, Massachusetts or ISO smoking mode;
[0035] b. Smoke capture: Use full smoke traps in series with absorption bottles or Cambridge filters, or Cambridge filters in series with absorption bottles, or use Cambridge filters or absorption bottles alone for e-cigarette smoke collection.
[0036] c. Configure standard series solutions: prepare series standard solutions with NNK, NNN, NAT and NAB standard substances; ①Weigh 10.0 mg NNK, NNN, NAT and NAB respectively, dissolve in methanol, and transfer to 4 10 mL brown volumetric flasks, methanol Constant volume; ②Respectively pipette 0.1 mL, 0.5 mL, 0.5 mL and 0.5 mL of the above solution into a 100 mL volumetric flask with methanol to a constant volume; ③ Pipette 10.0 mL ② of the solution into a 100 mL brown volumetric flask, with methanol to a constant volume; ④Pipette 0.1 mL, 0.2 mL, 0.5 mL, 1.0 mL, 2.0 mL, and 5.0 mL of the solution in ③ to 100 mL brown volumetric flasks, add 1 mL NAB-d4, NAT-d4, NNK with a concentration of 1.0 μg/mL, respectively -d4 and NNN-d4, use 0.1 mol/L ammonium acetate solution to make the volume constant to prepare a six-level series of standard solutions, the concentration of which is 10.0 ng/mL.
[0037] d. Preparation of test solution: Put the captured Cambridge filter and absorption solution into an Erlenmeyer flask, add ammonium acetate solution containing internal standard substance for extraction preparation, that is, the sample solution to be tested, the internal standard substance NNK-d4, NNN-d4, NAT-d4 and NAB-d4;
[0038] The preparation method of the ammonium acetate extraction solution containing the internal standard substances NNK-d4, NNN-d4, NAT-d4 and NAB-d4: ①Weigh 5.0 mg of the above four substances respectively, and dissolve the methanol in four 10 mL brown Volumetric flask and constant volume; ②Respectively transfer the 4 solutions in 1.0 mL ① to a 100 mL brown volumetric flask, and the volume of methanol is constant; ③ Pipette 10.0 mL ② the solution to a 100 mL brown volumetric flask, and the volume of methanol is constant; ④ Pipette 5.0 The solution in mL③ is transferred to a 500 mL volumetric flask, and the volume is constant with 0.1 mol/L ammonium acetate solution to prepare a 5.0 ng/mL ammonium acetate extraction solution containing internal standards.
[0039] e. Drawing the standard curve: absorb the prepared mixed standard solutions of different concentrations and inject them into the high performance liquid chromatography-triple quadrupole tandem mass spectrometry system to obtain the standard curves of the four substance concentrations and peak areas of NNK, NNN, NAT and NAB , Namely the one-variable linear regression equation;
[0040] f. Determination of sample liquid to be tested: Inject the prepared liquid to be tested into the high performance liquid chromatography-triple quadrupole tandem mass spectrometry system, and measure the ratio of the peak area of ​​the analyte to the internal standard, and substitute it into the unary linear regression equation to obtain the sample The content of the analyte in the test solution.
[0041] The liquid chromatography column is C18, and the liquid chromatography conditions:
[0042] Chromatographic column: C18 column (1.7 μm, 2.1×100 mm) or equivalent column, using Waters Acquity UPLC BEH C18 1.7 μm, 2.1×100 mm analytical column;
[0043] Flow rate: 0.1-1.0 mL/min;
[0044] Column temperature: 30-60 ℃;
[0045] Injection volume: 5-10 μL;
[0046] Mobile phase A: methanol, mobile phase B: 0.1-0.5% (V/V) acetic acid aqueous solution;
[0047] The gradient elution conditions are shown in the table below.
[0048] Fast gradient elution conditions for high performance liquid chromatography
[0049]
[0050] Or chromatographic column: C18 column (5 μm, 4.6×150 mm) or equivalent column; this project research uses Waters XTerra MS C18 5 μm 4.6×150 mm analytical column;
[0051] Flow rate: 0.1-1.0 mL/min;
[0052] Column temperature: 30-60 ℃;
[0053] Injection volume: 5-10 μL;
[0054] Mobile phase A: methanol, mobile phase B: 0.1-0.5% (V/V) acetic acid aqueous solution;
[0055] The gradient elution conditions are shown in the table below.
[0056] Ordinary high performance liquid chromatography gradient elution conditions
[0057]
[0058] In step f, the quantitative ion pairs selected by NNK, NNN, NAT, and NAB when the test solution is injected into the high performance liquid chromatography-triple quadrupole tandem mass spectrometry system are 208.1/122.1, 178.1/148.2, 190.1/160.1 and 192.1/162.1, respectively ; The qualitative transitions selected by NNK, NNN, NAT and NAB are 208.2/148.1, 178.2/120.01, 190.2/106.1, 192.1/133.1; NNK-d4, NNN-d4, NAT-d4 and NAB-d4 The quantitative ion pairs are 212.2/126.1, 182.1/152.2, 194.1/164.2, 196.2/166.2; the qualitative ion pairs selected by NNK-d4, NNN-d4, NAT-d4 and NAB-d4 are 212.2/110.1, 182.1/ 124.1, 194.1/110.1, 196.2/137.2.
[0059] The ion source of the high-performance liquid chromatography-triple quadrupole tandem mass spectrometry system described in step f is an electrospray ionization source (ESI); the scanning method is positive ion scanning; the detection method is multiple reaction monitoring (MRM); the electrospray voltage is 5500 V; ion source temperature is 650 ℃; auxiliary gas Gas1 pressure is 50.6 Psi; auxiliary gas Gas2 pressure is 70.0 Psi.
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