CN106840945B - A kind of flue gas dosimeter measuring method under full fume exposure experiment - Google Patents
A kind of flue gas dosimeter measuring method under full fume exposure experiment Download PDFInfo
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- 239000003546 flue gas Substances 0.000 title claims abstract description 80
- UGFAIRIUMAVXCW-UHFFFAOYSA-N Carbon monoxide Chemical compound [O+]#[C-] UGFAIRIUMAVXCW-UHFFFAOYSA-N 0.000 title claims abstract description 74
- 239000003517 fume Substances 0.000 title claims abstract description 53
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000002474 experimental method Methods 0.000 title claims abstract description 20
- 238000003380 quartz crystal microbalance Methods 0.000 claims abstract description 33
- 235000019504 cigarettes Nutrition 0.000 claims abstract description 19
- 239000002245 particle Substances 0.000 claims abstract description 17
- 239000000779 smoke Substances 0.000 claims abstract description 11
- 238000001514 detection method Methods 0.000 claims abstract description 5
- 239000003571 electronic cigarette Substances 0.000 claims abstract description 3
- 238000010438 heat treatment Methods 0.000 claims abstract 2
- 238000010790 dilution Methods 0.000 claims description 11
- 239000012895 dilution Substances 0.000 claims description 11
- SNICXCGAKADSCV-JTQLQIEISA-N (-)-Nicotine Chemical compound CN1CCC[C@H]1C1=CC=CN=C1 SNICXCGAKADSCV-JTQLQIEISA-N 0.000 claims description 10
- 229960002715 nicotine Drugs 0.000 claims description 10
- SNICXCGAKADSCV-UHFFFAOYSA-N nicotine Natural products CN1CCCC1C1=CC=CN=C1 SNICXCGAKADSCV-UHFFFAOYSA-N 0.000 claims description 10
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 9
- 230000004888 barrier function Effects 0.000 claims description 7
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- 239000007789 gas Substances 0.000 claims description 6
- NDJKXXJCMXVBJW-UHFFFAOYSA-N heptadecane Chemical compound CCCCCCCCCCCCCCCCC NDJKXXJCMXVBJW-UHFFFAOYSA-N 0.000 claims description 6
- 238000001819 mass spectrum Methods 0.000 claims description 5
- 230000000391 smoking effect Effects 0.000 claims description 5
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 230000008021 deposition Effects 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 238000003786 synthesis reaction Methods 0.000 claims description 4
- 238000012512 characterization method Methods 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 claims description 3
- 238000012544 monitoring process Methods 0.000 claims description 3
- 230000005540 biological transmission Effects 0.000 claims description 2
- 239000012159 carrier gas Substances 0.000 claims description 2
- 239000012930 cell culture fluid Substances 0.000 claims description 2
- 230000001186 cumulative effect Effects 0.000 claims description 2
- 239000003550 marker Substances 0.000 claims description 2
- 238000005070 sampling Methods 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 239000013078 crystal Substances 0.000 claims 1
- 238000001914 filtration Methods 0.000 claims 1
- 239000010453 quartz Substances 0.000 claims 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims 1
- 230000017105 transposition Effects 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 5
- 231100000750 In vitro toxicology Toxicity 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 4
- 238000012360 testing method Methods 0.000 abstract description 4
- 238000011156 evaluation Methods 0.000 abstract description 3
- 238000012502 risk assessment Methods 0.000 abstract description 3
- 235000019505 tobacco product Nutrition 0.000 abstract description 3
- 239000000443 aerosol Substances 0.000 abstract description 2
- 231100000580 in vitro toxicity testing Toxicity 0.000 abstract description 2
- 230000035945 sensitivity Effects 0.000 abstract description 2
- 230000008280 toxic mechanism Effects 0.000 abstract description 2
- 238000005259 measurement Methods 0.000 description 5
- 231100000419 toxicity Toxicity 0.000 description 3
- 230000001988 toxicity Effects 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000005374 membrane filtration Methods 0.000 description 2
- 230000007096 poisonous effect Effects 0.000 description 2
- 231100000820 toxicity test Toxicity 0.000 description 2
- 241000208125 Nicotiana Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- GSQIWDRAAXKDDL-UHFFFAOYSA-N heptadecane;methanol Chemical compound OC.CCCCCCCCCCCCCCCCC GSQIWDRAAXKDDL-UHFFFAOYSA-N 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012453 solvate Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000002110 toxicologic effect Effects 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 238000009602 toxicology test Methods 0.000 description 1
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N5/00—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid
- G01N5/02—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid by absorbing or adsorbing components of a material and determining change of weight of the adsorbent, e.g. determining moisture content
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/06—Investigating concentration of particle suspensions
- G01N15/0606—Investigating concentration of particle suspensions by collecting particles on a support
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Dispersion Chemistry (AREA)
- Sampling And Sample Adjustment (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
A kind of flue gas dosimeter measuring method detected under full fume exposure experiment carries out real-time, the quantitative determination of flue gas dosage in full fume exposure storehouse using quartz crystal microbalance chip in conjunction with plug-in type Tissue Culture Dish；Maximum feature of the invention is: the practical microenvironment of simulated flue gas exposure, in real time, sensitive, the full fume exposure storehouse inner wrap strip flue gas of quantitative detection actual concentrations (mass concentration and nicotinic density including flue gas particles), the dose-effect relationship of flue gas toxity effect is accurately characterized, provides true flue gas dosage basis for cigarette smoke in vitro toxicology Effect Evaluation.The toxic mechanism of cigarette smoke can be preferably explained using the method for the present invention, be that the health risk assessment of tobacco product studies providing method basis and technical support.Meanwhile the in vitro toxicity testing research of the aerosol of this method applicability is wide, can be applied to cigarette, electronic cigarette, heating are not burnt cigarette and separate sources, test result is reproducible, and stability is high, and sensitivity is strong.
Description
Technical field
The present invention relates to the research of the in vitro toxicology of cigarette smoke, the specifically full fume exposure experiments of a kind of detection
Under flue gas dosimeter measuring method.
Background technique
Cigarette smoke in vitro toxicology research based on full fume exposure is that smoke of tobacco in vitro toxicity is evaluated in recent years
One of hot spot.Carrying out full fume exposure experiment can be more comprehensively true to reflect volume with the internal microenvironment of simulated flue gas exposure
The biological effect of cigarette flue gas.Toxicologic study will be based on accurate proof load, the real-time monitoring of flue gas dosage and precisely control
System is to influence a key factor of full fume exposure experimental result reliability.Currently, when carrying out the experiment of full fume exposure, cruelly
Practical flue gas concentration in dew storehouse is difficult to measure, such as dilute with diluent air flow velocity, flue gas usually by the way of characterizing indirectly
Release the forms characterizations such as ratio, suction cigarette quantity.Although the characterizing method of these flue gas dosage may be implemented between test sample
The purpose that Difference In Toxicity compares, but for the true flue gas exposure concentrations under physical condition and be unaware of.Therefore, a kind of reality is established
The measuring method of flue gas dosage under the conditions of border, actual, real-time flue gas concentration in the full fume exposure storehouse of exact representation, for cigarette
The in vitro toxicity evaluation of straw-made articles is of great significance, and more accurately reflects the cigarette smoke biology under true flue gas concentration
Dose-effect relationship.
Summary of the invention
The purpose of the present invention is based on above-mentioned condition and the flue gas agent under a kind of full fume exposure experiment of detection for providing
Amount learns measuring method.Using the method for foundation full fume exposure can be tested in fume exposure storehouse flue gas concentration (including
Flue gas particles mass concentration and nicotinic density) real-time monitoring, quantitatively characterizing are carried out, it is provided accurately for toxicology test result
Poisonous effect dosage basis.Providing method basis is studied for the health risk assessment of tobacco product.
The invention mechanism of this method foundation is: using quartz crystal microbalance (Quartz Crystal
Microbalance, QCM) chip and plug-in type Tissue Culture Dish (Transwell) combine, carry out cigarette in full fume exposure storehouse
Real-time, the quantitative determination of gas dosage.QCM chip can obtain the actual concentrations of flue gas particles in exposed storehouse, be toxicity test
Test result provide dosimetric data support, the bio-toxicity effect of flue gas is more accurately described.
The purpose of the present invention is be achieved through the following technical solutions:
A kind of flue gas dosimeter measuring method detected under full fume exposure experiment, uses quartz crystal microbalance (Quartz
Crystal Microbalance, QCM) chip and plug-in type Tissue Culture Dish (Transwell) combine, it is sudden and violent to carry out full flue gas
Reveal real-time, the quantitative determination of flue gas dosage in storehouse；Specific step is as follows:
1) full fume exposure experiment
Plug-in type Tissue Culture Dish is placed in full fume exposure storehouse, quartz crystal microbalance chip is placed in
On the microporous barrier of the cell Transwell, fume exposure experiment, full fume exposure storehouse and smoking machine and flue gas dilution system phase are carried out
Connection, the fresh smoke that smoking machine suction generates is after flue gas dilution system is mixed with synthesis of air, with the stream of 5 mL/min
Speed enters in full fume exposure storehouse, the dilution flue gas real-time contact in QCM wafer surface and exposure storehouse；The cell Transwell
Microporous barrier under cell culture fluid can be perfused, to simulate the microenvironment of actual exposed；
2) flue gas dosimetry-flue gas particles content assaying
QCM chip in full fume exposure storehouse can be taken off after fume exposure, carry out manual weighing, and measurement QCM is brilliant
The quality of the flue gas particles of piece surface deposition, QCM chip carry out the quality difference value after full fume exposure divided by QCM wafer surface
The fume exposure cumulative concentration (the flue gas particle amount of substance of per surface area) in exposed storehouse can be obtained in product；When QCM chip with
When current signal sensor connects, current signal sensor can be converted vibration of the QCM chip under flue gas particles deposition
The real-time of flue gas dosage, automatic ration prison are realized to characterize the real-time mass concentration variation of flue gas in exposed storehouse for electric signal
It surveys；
3) flue gas dosimetry-smoking nicotine concentration mensuration
Using the nicotine in flue gas as the chemical marker of characterization flue gas dosage, the flue gas deposited with QCM wafer surface
The nicotine quality of grain object carrier band is come the flue gas dosage that reflects in full fume exposure storehouse；
After full fume exposure experiment, the QCM chip on the cell Transwell microporous barrier in exposure storehouse is taken out, is turned
It is placed in conical flask, is added and extracts solution, 30 min-60 min are vibrated under 180 rpm, after 0.22 μm of membrane filtration,
Solution is added in chromatogram bottle after drawing 1 mL filter, carries out gas chromatography-mass spectrum (GC-MS) analysis, quantitative determines to nicotine.
The extraction solution is isopropanol or methanol.
Maximum feature of the invention is: the practical microenvironment of simulated flue gas exposure, real-time, sensitive, the full flue gas of quantitative detection
The actual concentrations (mass concentration and nicotinic density including flue gas particles) of exposure storehouse inner wrap strip flue gas, accurately characterize flue gas
The dose-effect relationship of poisonous effect provides true flue gas dosage basis for cigarette smoke in vitro toxicology Effect Evaluation.It answers
The toxic mechanism of cigarette smoke can be preferably explained with the method for foundation, provided for the health risk assessment research of tobacco product
Method basis and technical support.Meanwhile the method applicability of foundation is wide, can be applied to cigarette, electronic cigarette, heats the cigarette that do not burn
And the in vitro toxicity testing research of the aerosol of separate sources, test result is reproducible, and stability is high, and sensitivity is strong.
Detailed description of the invention
(n=3, n are the parallel independent measurement number of setting to flue gas dosimeter measurement result, i.e., under the full fume exposure of Fig. 1
Have the measurement result in 3 parallel exposure storehouses under one dosage),
Wherein: Figure 1A is flue gas particles mass concentration (the μ g/ of different dilution ratio flue gases in full fume exposure storehouse
cm2), Figure 1B is the nicotinic density (μ g/mL) of the flue gas particles carrier band of different dilution ratio flue gases in full fume exposure storehouse.
Specific embodiment
The present invention is described further below in conjunction with example (attached drawing):
Choose the flue gas dosimeter measurement experiment that reference cigarette 3R4F carries out full fume exposure.By plug-in type Tissue Culture Dish
(Transwell) be placed in full fume exposure storehouse, by quartz crystal microbalance (Quartz Crystal Microbalance,
QCM) chip is placed on the microporous barrier of the cell Transwell, carries out fume exposure experiment.Full fume exposure storehouse and smoking machine and
Flue gas dilution system is connected, and smoking machine aspirates 3R4F reference cigarette, the fresh smoke of generation according to ISO condition (35/2/60)
It after being mixed with synthesis of air different in flow rate, is entered in full fume exposure storehouse with the flow velocity of 5 mL/min, synthesis of air setting 5
A flow velocity, respectively 0.25,0.5,1.0,2.0,4.0 L/min.QCM wafer current signal transducer is opened, full flue gas is recorded
Flue gas particles mass concentration in exposure storehouse.After full fume exposure experiment, by the cell Transwell micropore in exposure storehouse
QCM chip on film takes out, and is placed in conical flask, and 10 mL methanol solvates (or isopropanol solvent) are added, and is added 500 μ L's
Internal standard n-heptadecane methanol solution (n-heptadecane concentration is 1 mg/mL)；30 min are vibrated under 180 rpm, and 1.5 mL are sucked out
Extract liquor is drawn 1 mL solution and is added in chromatogram bottle, carry out gas chromatography-mass spectrum (GC-MS) after 0.22 μm of membrane filtration
Analysis.GC-MS analysis condition is as follows:
--- injector temperature: 260 DEG C；
--- sampling volume: 2 μ L；
--- split ratio: 10:1；
--- carrier gas: He gas, flow velocity: 1.0 mL/min；
--- transmission line temperature: 280 DEG C；
--- chromatographic column: DB-5MS (0.25 μm of the μ m of 30 m × 250) capillary chromatographic column；
--- temperature programming condition:, keeping 3 min by 100 DEG C of initial temperature, rises to 260 DEG C with 8 DEG C/min, keeps 5
min。
--- mass spectrum ionizes mode: EI ionization, 70 eV；
--- scan pattern: SIM mode, nicotine (m/z 84), n-heptadecane (m/z 57).
Experimental result is as shown in Figure 1, Figure 1A is the flue gas particle substance of different dilution ratio flue gases in full fume exposure storehouse
Measure concentration (μ g/cm2), Figure 1B is the nicotinic density of the flue gas particles carrier band of different dilution ratio flue gases in full fume exposure storehouse
(μ g/mL) (quality of nicotine in i.e. every milliliter extraction solution).
Claims (2)
1. a kind of flue gas dosimeter measuring method detected under full fume exposure experiment, it is characterised in that: use Quartz Crystal day
Flat QCM chip carries out the real-time, fixed of flue gas dosage in full fume exposure storehouse in conjunction with plug-in type Tissue Culture Dish Transwell
It is fixed to measure；Specific step is as follows:
1) full fume exposure experiment
Plug-in type Tissue Culture Dish is placed in full fume exposure storehouse, quartz crystal microbalance chip is placed in Transwell
On the microporous barrier of cell, fume exposure experiment is carried out, full fume exposure storehouse is connected with smoking machine and flue gas dilution system, smokes
The fresh smoke that machine suction generates is entered entirely after flue gas dilution system is mixed with synthesis of air with the flow velocity of 5 mL/min
Dilution flue gas real-time contact in fume exposure storehouse, in QCM wafer surface and exposure storehouse；Under the microporous barrier of the cell Transwell
Cell culture fluid can be perfused, to simulate the microenvironment of actual exposed；
2) flue gas dosimetry-flue gas particles content assaying
QCM chip in full fume exposure storehouse can be taken off after fume exposure, carry out manual weighing, measure QCM chip table
The quality of the flue gas particles of face deposition, QCM chip carry out the quality difference value after full fume exposure divided by QCM wafer surface product i.e.
It can obtain exposing the fume exposure cumulative concentration in storehouse；When QCM chip is connect with current signal sensor, current signal sensing
Vibration of the QCM chip under flue gas particles deposition can be converted to electric signal by device, to characterize the reality of flue gas in exposed storehouse
When mass concentration change, realize real-time, the automatic ration monitoring of flue gas dosage；
3) flue gas dosimetry-smoking nicotine concentration mensuration
Using the nicotine in flue gas as the chemical marker of characterization flue gas dosage, the flue gas particles deposited with QCM wafer surface
The nicotinic density of carrier band is come the flue gas dosage that reflects in full fume exposure storehouse；
After full fume exposure experiment, the QCM chip in exposure storehouse on the cell Transwell microporous barrier is taken out, transposition in
In conical flask, it is added and extracts solution isopropanol or methanol, 30 min-60 min are vibrated under 180 rpm, through 0.22 μm of filter
After film filtering, solution is added in chromatogram bottle after drawing 1 mL filter, is carried out gas chromatography-mass spectrum GC-MS analysis, is determined nicotine
It is fixed to measure, and GC-MS analysis condition is as follows:
Injector temperature: 260 DEG C；Sampling volume: 2 μ L；Split ratio: 10:1；Carrier gas: He gas, flow velocity: 1.0 mL/min；Transmission
Line temperature: 280 DEG C；Chromatographic column: DB-5MS capillary chromatographic column, 0.25 μm of μ m of specification 30 m × 250；Temperature programming item
Part:, keeping 3 min by 100 DEG C of initial temperature, rises to 260 DEG C with 8 DEG C/min, keeps 5 min；Mass spectrum ionizes mode: EI ionization,
70 eV；Scan pattern: SIM mode, nicotine m/z 84, n-heptadecane m/z 57.
2. the flue gas dosimeter measuring method under the full fume exposure experiment of detection according to claim 1, it is characterised in that:
The lower flue gas dosimeter survey of the full fume exposure experiment of the measuring method is suitable for cigarette, electronic cigarette, heating are not burnt cigarette
It is fixed.
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| CN109557254B (en) * | 2018-11-14 | 2021-08-03 | 云南中烟工业有限责任公司 | Method for establishing standardized cigarette smoke mouse mouth and nose exposure method |
| CN109557005B (en) * | 2018-11-14 | 2021-09-14 | 云南中烟工业有限责任公司 | Cigarette smoke mouse mouth and nose exposure method |
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