CN102353741A - Method for measuring contents of four kinds of tobacco-specific nitrosamines in cigarette mainstream smoke - Google Patents
Method for measuring contents of four kinds of tobacco-specific nitrosamines in cigarette mainstream smoke Download PDFInfo
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Abstract
The invention discloses a method for measuring contents of four kinds of tobacco-specific nitrosamines in a cigarette mainstream smoke. The method comprises the following steps of: utilizing a glass fiber filter disc to capture tobacco-specific nitrosamines in the cigarette mainstream smoke and extracting the tobacco-specific nitrosamines by using a water solution containing interior labels, methanol and ethanol; and filtering an extraction solution and directly utilizing a high performance liquid chromatography-mass spectrometer (LC-MS/MS) to detect the contents of the four types of the tobacco-specific nitrosamines (TSNAs) in the cigarette mainstream smoke. According to the detection method provided by the invention, new liquid chromatography, fluid phase and gradient elution procedures are adopted to completely separate the four types of the tobacco-specific nitrosamines (TSNAs), so that the selectivity, resolution ratio and sensitivity of chromatography separation are improved. The method provided by the invention has the advantages of simple pretreatment, good separation effects, short sample analysis time, high detection sensitivity and the like; and rapid and accurate detection on the tobacco-specific nitrosamines (TSNAs) in the cigarette mainstream smoke is realized.
Description
Technical field
The present invention relates in a kind of cigarette mainstream flue gas four unique nitrosamine (TSNAs) the Determination on content methods that grow tobacco, belong to tobacco and technical field of chemistry.
Background technology
Flue gas is a kind of very complicated potpourri; But has only 0.6% objectionable impurities; It is carcinogenic and suspect carcinogen that 0.2% (kind more than 60) wherein only arranged, and the material that is piped off by IRAC (international cancer research association) has seven types, wherein just comprises tobacco-specific nitrosamine (TSNAs).Also have in most of physical environment air in the flue gas,, in the coffee of urban air, vehicle exhaust, kitchen fume, grilled beef, baking and fruit, vegetables and cereal products, discovery is arranged all like benzo [a] pyrene (B[a] P).But tobacco-specific nitrosamine (TSNAs) is an exception; Be one type of carcinogen that in tobacco solarization system, processing, fermentation and combustion process, produces, all identified these materials at tobacco product, flue gas, environment flue gas (ETS), smokeless tobacco and other tobacco product such as cigar and India cigar the inside.
Found in the world that at present more than 300 kind of N-nitrosamine has carcinogenicity to more than 40 kinds of animals, but known in tobacco product and the flue gas have only eight kinds, wherein just comprise four unique nitrosamines (TSNAs) that grow tobacco: NNN, NNK, NAT, NAB.Generally believe that at present TSNAs can make the people suffer from lung cancer, particularly NNN and NNK.No matter show with rodent research, be feeding or injection, and NNK all can cause lung tumors-epidermal cell benign tumour, and adenocarcinoma of lung-malignant epithelioma.Research is illustrated in the metabolic processes; NNK can make dna methylation (from this tissue, having isolated the O6-methyl guanine) in living animal or the stripped tissue; Molecular biologist thinks that in the cytogenetics password, having the O6-methyl guanine is a kind of chemical damage to DNA, might be carcinogenic.The zooscopy test report of IRAC thinks that then NNN also is that a meeting causes multiple carcinogenic danger composition.In addition, NAS estimates, the smoker who takes out A box of cigarette every day is 17 milligrams to the contact amount of TSNAs.The route of exposure of TSNAs has respiratory system suction, alimentary canal absorption to contact with skin.Smoking or contact environment flue gas all can touch TSNAs.Carcinogenicity TSNAs does not all have secure threshold.
The mechanism of production of TSNAs: generally believe that the mechanism that TSNAs produces in the tobacco product is the natural nitrosification of nicotiana alkaloids.TSNAs and content of nitrite are atomic in the fresh tobacco, it is generally acknowledged that the formation of TSNAs and accumulation are to occur in to pluck afterwards and the modulation treatment stage.A kind of hypothesis is arranged recently, say so because this in stage microorganism be nitrite and other nitrogen-containing oxide (NOx compound) with nitrate reduction, form TSNAs with the nicotiana alkaloids effect then.Dialogue rib tobacco product also has similar hypothesis.But for the TSNAs mechanism of production of baking type tobacco different hypothesis is in addition arranged also, the gas of naked light combustion furnace discharging has directly got into barn, the NOx in the gas spontaneously with tobacco in alkaloid react and form TSNAs.
Present existing tobacco-specific nitrosamine analyzing detecting method
1) vapor-phase chromatography: content of nitrosamines is atomic in the cigarette mainstream flue gas; Exist system extremely complicated; Therefore sample is carried out before conventional gas chromatography (GC) analyzes, for reducing that analysis background disturbs and, passing through steps such as extract and separate, enrichment and purifying earlier to the pollution of chromatographic column.Extractant commonly used has: citrate buffer solution, diluted sodium hydroxide solution, citrate buffer solution one toluene mixed solution etc.The exemplary steps of extraction, purifying TSNAs is: have the glass fibre filter disc of main flume granule phase substance to put in people's conical flask capture, add citrate buffer solution, place ultrasonic Extraction in the ultrasonic generator, use dichloromethane extraction again; Extract filters behind anhydrous sodium sulfate drying; The round-bottomed flask that fills filtrating is connected Rotary Evaporators; Logical high pure nitrogen rotary evaporation; Be concentrated into about 5mL under 40 ℃, alkali alumina and concentrate sample behind 110 ℃ of following activation 2h added chromatographic column, with methylene chloride one acetone elution chromatography post; Collect whole eluents; After marking liquid in eluent, accurately adding, be concentrated into about 1mL, the sample introduction analysis under 40 ℃.
2) gas chromatography one heat energy detecting device method: vapor-phase chromatographies in 1973 just have been used to the analysis of TSNAs; But because the content of each TSNAs has nothing in common with each other; NNN wherein; The content of NAT and NNK is relatively large; The content of NAB is atomic, and early stage its precision of vapor detection device is difficult to satisfy the requirement of quantitative test.In 1975, thermal energy analyzer (TEA) was used to TSNAs is detected, and TEA is a kind of vapor detection technology to volatilization and non-volatile N one nitrosamino group compound trace analysis.After this coupling of TEA and GC technology has improved selectivity and the sensitivity to the N-nitroso compound to a certain extent.In recent years, the someone adopts gas chromatograph to be equipped with nitrogen phosphorous detector and measures TSNAs, has obtained effect preferably, but not very good to the atomic NAB testing result of content.
3) LC-MS/MS method: can measure the TSNAs content in multiple pipe tobacco and the mainstream smoke simultaneously in fast quantification ground through utilizing liquid chromatography-mass spectrography/mass spectrometry method.Its key step is: in the cambridge filter of offal or collection flue gas, add the ammonium acetate solution of 0.1 mol and the internal standard compound of deuterate, shaking out 30min filters, and filtrating is used for LC-MS/MS to be analyzed, and only be about 8min analysis time.This method has following characteristics: 1. quantity of sample handling is little, and step is simple, does not need purifying, and the processing time is short; 2. detectability is lower, highly sensitive than gas chromatography and mass spectromentry coupling method; 3. lack analysis time, the sample analysis amount is big; 4) stable performance, favorable reproducibility.But present LC-MS/MS method since the liquid-phase chromatographic column that adopts exist four grow tobacco that unique nitrosamine (TSNAs) separating effect is undesirable, the moving phase used and gradient elution cause analyzing the peak and send out fork, seriously trail and shortcoming such as noise is higher.
Therefore research set up a kind of fast, accurately, four unique nitrosamine (TSNAs) content detecting method that grow tobacco in the good separating effect, highly sensitive, cigarette mainstream flue gas that reappearance is good, remain a current urgent task of tobacco business.
Summary of the invention
The purpose of this invention is to provide that a kind of pre-treatment is simple, four unique nitrosamine (TSNAs) the Determination on content methods that grow tobacco in the good separating effect, sample analysis time is short, detection accuracy is high cigarette mainstream flue gas.
The four unique nitrosamine Determination on content methods that grow tobacco in a kind of cigarette mainstream flue gas: utilize the glass fibre filter disc to capture cigarette mainstream flue gas; With the aqueous solution extraction that contains interior mark, methyl alcohol and ammonium acetate; Wherein the volume ratio of methyl alcohol and water is 0-1: 15-20, and the ammonium acetate concentration of adding is the 0.08-0.12 mol; The concentration of four kinds of internal standard compounds respectively is 3-10ng/mL; Extract directly utilizes high performance liquid chromatography-mass spectrometer that four content that grow tobacco unique nitrosamine in the cigarette mainstream flue gas are detected after filtering; Said four unique nitrosamines that grow tobacco are N-nitroso-nornicotine (NNN), N-nitroso-anabasine (NAB), N-nitroso-anatabine (NAT), 4-(N-methyl-N-nitrosamine)-1-(3-pyridine radicals)-butanone (NNK);
The liquid-phase chromatographic column that said high performance liquid chromatograph adopted is an Atlantis dC18 chromatographic column, and other parameter of said high performance liquid chromatograph is:
Sample size: 1-10 microlitre;
Flow velocity: 0.1-0.3ml/min;
Column temperature: 30-70 ℃;
Mobile phase A is the ammonium acetate methanol aqueous solution, and wherein the volume ratio of methyl alcohol and water is 0-1: 15-20, ammonium acetate concentration be the 3-8 mM/liter; Mobile phase B is a methyl alcohol;
The gradient elution program: the 0-3min eluent is the A of volumn concentration 85%-95% and the B of 5-15%; The B of the A:40-60% of 3-8min40%-60%; The A of the above 85%-95% of 8min and the B of 5-15%;
Said mass spectrometric detected parameters is:
Ionization mode: electron spray (ESI);
Scan pattern: positive ion;
Atomization gas Nebulizer Gas (NEB): 7.00-9.00;
Gas curtain gas Curtain Gas (CUR): 7.00-9.00;
Collision gas Collision Gas (CAD): 6.00-8.00;
Ion gun injection electric IonSpray Voltage (IS): 5000.00-5500.00;
Ion source temperature Temperature (TEM): 380.00-420.00.
The said four unique nitrosamine internal standard compound standard items that grow tobacco are four deuterium generation-N-nitroso-nornicotine (d
4-NNN), four deuterium generation-N-nitroso-anabasine (d
4-NAB), four deuterium generation-N-nitroso-anatabine (d
4-NAT), four deuterium generation-4-(N-methyl-N-nitrosamine)-1-(3-pyridine radicals)-butanone (d
4-NNK).
Said four grow tobacco unique nitrosamine measured objects and the detected parameters of internal standard compound is following separately:
1. the preparation of standard reserving solution: accurately take by weighing the four unique nitrosamine standard items that grow tobacco respectively, mix, be mixed with concentration and respectively be the standard reserving solution of 0.8-1.2mg/mL with dissolve with methanol, constant volume;
2. the preparation of mark storing solution in: accurately take by weighing four respectively and grow tobacco the unique nitrosamine deuterium for the internal standard compound standard items, mix and use dissolve with methanol, constant volume is mixed with concentration and respectively is the interior mark storing solution of 0.8-1.2mg/mL;
The series standard solution of 7-8 variable concentrations gradient will be formulated as during detection after the standard reserving solution dilution; Concentration is in the 0-100ng/mL scope.
Cigarette mainstream flue gas sample pre-treatments: have the glass fibre filter disc of cigarette sample main flume to be positioned in the conical flask of 50mL ground band plug capture; Add the 10-15mL extract; Get the 0.8-1.2mL extract behind the mechanical oscillation 20-40min, adopt usefulness to be analyzed behind the 0.22 μ m water membrane filtration.
Utilize the high performance liquid chromatography-tandem mass appearance that series standard solution is detected respectively; Four the grow tobacco chromatographic peak area and the corresponding internal standard compound peak area ratios of unique nitrosamine that obtain are right; Draw respectively four grow tobacco unique nitrosamine typical curve and obtain regression equation; Under the same conditions sample is detected then; To be measured four chromatographic peak areas that grow tobacco unique nitrosamine that obtain are brought in the corresponding regression equation, can be obtained in the sample corresponding four content that grow tobacco unique nitrosamine.
Step By Condition to the suction of cigarette when said cigarette mainstream flue gas is collected is: the cigarette of choosing is propped up and glass fibre filter disc balance 48 hours under 22 ± 1 ℃, relative humidity 60 ± 2% conditions; Under this condition, use smoking machine that cigarette is aspirated then; Capture cigarette smoke with the glass fibre filter disc; Every mouthful was aspirated 2 seconds; Draw volume 35mL; Every mouthful of interval 58 seconds, every group of suction 4-5 props up cigarette.
Assay method provided by the invention has overcome shortcomings such as the pre-treatment step that exists in existing general high performance liquid chromatography-thermal energy analyzer (LC-TEA) detection method is loaded down with trivial details, each sample analysis time is long, detection sensitivity is low.
Present existing LC-MS/MS assay method exists and four grows tobacco that unique nitrosamine (TSNAs) separating effect is undesirable, fork is sent out at the sample analysis peak, seriously trails and shortcoming such as noise is higher; The present invention has adopted new liquid-phase chromatographic column, moving phase and gradient elution program; Make four unique nitrosamines (TSNAs) that grow tobacco obtain fully separating therefore selectivity, resolution and the sensitivity that has improved chromatographic resolution.
Description of drawings
Fig. 1 is the chromatogram of standard solution.
Fig. 2 is the standard solution chromatogram of (containing internal standard compound).
Fig. 3 is the four unique nitrosamine chromatograms of (containing internal standard compound) that grow tobacco in certain trade mark cigarette mainstream flue gas.
Mark among the figure, 1, NNN, 2, NNK, 3, NAT, 4, NAB, 5, (d
4-NNN), 6, (d
4-NNK), 7, (d
4-NAT), 8, (d
4-NAB).
Embodiment
Below four unique nitrosamine (TSNAs) the Determination on content methods that grow tobacco in the cigarette mainstream flue gas of the present invention are further illustrated, and can not limit the present invention.
Embodiment 1
1, instrument and reagent
The API of U.S. Applied Biosystems company 3000 type triple quadrupole bar tandem mass spectrometers are furnished with electric spray ion source (Turbo IonSpray) and Analyst 1.4 system softwares; U.S. Agilent company 1100 type high performance liquid chromatographs, band automatic sampler and constant temperature oven; Britain Filtrona company 450 types, 20 passage smoking machines; Electronic balance (sensibility reciprocal 0.1mg), adjustable mechnical oscillator circles round.
Methyl alcohol (chromatographically pure), ammonium acetate (chromatographically pure), deionized water; Four standard items that grow tobacco unique nitrosamine NNN, NNK, NAB, NAT, purity are all>=98%; Four kinds of corresponding deuteriums are for internal standard compound d
4-NNN, d
4-NNK, d
4-NAB, d
4The standard items of-NAT, purity are all>=99%.
2, the preparation of solution
The preparation of standard reserving solution: in the 50mL beaker, accurately take by weighing NNN, NNK, NAB, NAT standard items about about 100mg respectively; Accurately to 0.0001g; Add after about 30mL methyl alcohol dissolves fully; Be transferred in the 100mL volumetric flask; Wash the beaker inwall 3-5 time with small amount of methanol; Cleansing solution together is transferred in the 100mL volumetric flask, at last with methanol constant volume to scale.
The preparation of interior mark storing solution: in the 50mL beaker, accurately take by weighing the d about about 100mg respectively
4-NNN, d
4-NNK, d
4-NAB, d
4-NAT standard items accurately to 0.0001g, add after about 30mL methyl alcohol dissolves fully, are transferred in the 100mL volumetric flask, and with small amount of methanol washing beaker inwall 3-5 time, cleansing solution together is transferred in the 100mL volumetric flask, at last with methanol constant volume to scale.
Filter disc extract: in 950mL water and 50mL methyl alcohol mixed liquor, add in 7.7g ammonium acetate and the 5 μ L and mark storing solution, fully dissolving back mixing; The filter disc extract is promptly joined promptly and is used.
The preparation of primary standard solution: accurately pipette 50 μ L standard reserving solutions to the 100mL volumetric flask, be diluted to scale with filter disc extract constant volume.This solution must promptly be joined promptly and used.
The preparation of series standard solution: accurately pipette 0.02mL, 0.1mL, 0.5mL, 1.0mL, 2.0mL, 5.0mL, 10.0mL primary standard solution respectively to the 50mL volumetric flask; Be diluted to the standard solution that scale obtains 7 different series concentration with filter disc extract constant volume; Successively this series standard solution and filter disc extract are carried out the LC-MS/MS analyzing and testing, the production standard curve.
3, the collection of sample and pre-treatment
The suction of cigarette: with cigarette balance 48 hours under 22 ± 1 ℃, relative humidity 60 ± 2% conditions, under this condition, use smoking machine that cigarette is aspirated then, the main flume of suction is captured through the glass fibre filter disc.Every mouthful of suction 2 seconds, draw volume 35mL, every mouthful of interval 58 seconds, every group of suction 4-5 props up cigarette.
Sample extraction: take out and capture the glass fibre filter disc that the main flume granule phase substance is arranged; Be transferred in the 50mL conical flask; Accurately add 10mL filter disc extract; Get an amount of (about 0.8-1.2mL) extract behind the mechanical oscillation 30min behind 0.22 μ m water membrane filtration; Move on in the 2mL chromatogram bottle, can carry out LC-MS/MS and analyze.
4, detect
The high performance liquid chromatograph condition:
Liquid-phase chromatographic column: Waters Atlantis dC18 3 μ m 2.1 * 150mm Column.
Sample size: 5 microlitres;
Flow velocity: 0.2ml/min;
Column temperature: 60 ℃;
Mobile phase A: in 950mL water and 50mL methyl alcohol mixed liquor, add the 0.4g ammonium acetate, fully dissolving back mixing.
Mobile phase B is a methyl alcohol,
The gradient elution program: the 0-3min eluent is the A of volumn concentration 90% and 10% B; The A of 3-8min50% and 50% B; The A of 8-12min above 90% and 10% B;
The tandem mass spectrometer conditional parameter:
Ionization mode: electron spray (ESI);
Scan pattern: positive ion;
Atomization gas Nebulizer Gas (NEB): 8.00;
Gas curtain gas Curtain Gas (CUR): 8.00;
Collision gas Collision Gas (CAD): 7.00;
Ion gun injection electric IonSpray Voltage (IS): 5500.00;
Ion source temperature Temperature (TEM): 400.00;
Said four grow tobacco unique nitrosamine (TSNAs) measured objects and the main detected parameters of internal standard compound is following separately.
Analyzing and testing step: at first utilize high performance liquid chromatography-mass spectrometer (LC-MS/MS) that series standard solution is detected respectively; Four grow tobacco chromatographic peak area and the corresponding internal standard compound peak area ratio of unique nitrosamine (TSNAs) with what obtain to drawing four calibration curves that grow tobacco unique nitrosamine (TSNAs) respectively; Calculate the regression equation of calibration curve, four the grow tobacco testing results of unique nitrosamine (TSNAs) standard model are as shown in table 1 below.
The grow tobacco testing result of unique nitrosamine (TSNAs) standard model of table 1, four
The detection of cigarette sample: under the same conditions sample is detected; Four chromatographic peak areas that grow tobacco unique nitrosamine (TSNAs) in the sample are brought in the corresponding regression equation, can be obtained in the sample corresponding four content that grow tobacco unique nitrosamine (TSNAs).
Fig. 1 is the chromatogram of standard solution.From figure, can find out, adopt detection method of the present invention that four unique nitrosamines (TSNAs) that grow tobacco are had good separating effect.
Fig. 2 is the standard solution chromatogram of (containing internal standard compound).
Fig. 3 is the chromatogram of certain trade mark cigarette sample, also can find out that from figure four unique nitrosamines that grow tobacco have all realized baseline separation, has improved the accuracy of detection by quantitative.
5, the precision of assay method of the present invention (n=5), the recovery and detectability are as shown in table 2.
The precision of table 2. method, the recovery and detectability result
6, the calculating of content
The four mensuration results that grow tobacco unique nitrosamine (TSNAs) in every cigarette mainstream flue gas, calculate according to formula:
q＝S×V/n
In the formula:
Q is the detected value of tobacco-specific nitrosamine in every cigarette mainstream flue gas (TSNAs) content, and unit is every of nanogram (ng/cig);
S is the detectable concentration of tobacco-specific nitrosamine (TSNAs) in the cigarette mainstream flue gas sample extraction liquid, and unit is every milliliter of nanogram (ng/mL);
V is cigarette mainstream flue gas sample extraction liquid long-pending (mL);
N is the number of each test smoking cigarette, and unit is (cig).
Adopt the assay method in the embodiment, test cigarette sample A, B, C, D are detected, test specimen information and conventional index thereof are as shown in table 3:
Table 3 test specimen information and conventional index thereof detect data
Four unique nitrosamine (TSNAs) Determination on content that grow tobacco adopt the inventive method in the test specimen cigarette mainstream flue gas, and it is as shown in table 4 to measure the result:
| Numbering | The cigarette type | NNN (ng/ props up) | NNK (ng/ props up) | NAT (ng/ props up) | NAB (ng/ props up) | Total amount (ng/ props up) |
| A | Fire-cured tobacco type | 2.55 | 3.60 | 5.85 | 0.60 | 12.60 |
| B | Fire-cured tobacco type | 3.08 | 5.11 | 6.32 | 0.75 | 15.26 |
| C | Mixed type | 68.31 | 34.20 | 66.81 | 6.25 | 175.57 |
| D | Mixed type | 104.79 | 50.40 | 103.87 | 10.94 | 270.00 |
Claims (10)
1. four unique nitrosamine Determination on content methods that grow tobacco in the cigarette mainstream flue gas; It is characterized in that: utilize the glass fibre filter disc to capture cigarette mainstream flue gas; With the aqueous solution extraction that contains interior mark, methyl alcohol and ammonium acetate; Wherein the volume ratio of methyl alcohol and water is 0-1: 15-20, and the ammonium acetate concentration of adding is the 0.08-0.12 mol; The concentration of four kinds of internal standard compounds respectively is 3-10ng/mL; Extract directly utilizes high performance liquid chromatography-mass spectrometer that four content that grow tobacco unique nitrosamine in the cigarette mainstream flue gas are detected after filtering; Said four unique nitrosamines that grow tobacco comprise N-nitroso-nornicotine, N-nitroso-anabasine, N-nitroso-anatabine, 4-(N-methyl-N-nitrosamine)-1-(3-pyridine radicals)-butanone.
The liquid-phase chromatographic column that said high performance liquid chromatograph adopted is an Atlantis dC18 chromatographic column, and other parameter of said high performance liquid chromatograph is:
Sample size: 1-10 microlitre;
Flow velocity: 0.1-0.3ml/min;
Column temperature: 30-70 ℃;
Mobile phase A is the ammonium acetate methanol aqueous solution, and wherein the volume ratio of methyl alcohol and water is 0-1: 15-20, ammonium acetate concentration be the 3-8 mM/liter; Mobile phase B is a methyl alcohol;
The gradient elution program: the 0-3min eluent is the A of volumn concentration 85%-95% and the B of 5-15%; The B of the A:40-60% of 3-8min40%-60%; The A of the above 85%-95% of 8min and the B of 5-15%.
2. assay method according to claim 1 is characterized in that:
Said mass spectrometric detected parameters is:
Ionization mode: electron spray;
Scan pattern: positive ion;
Atomization gas: 7.00-9.00;
Gas curtain gas: 7.00-9.00;
Collision gas: 6.00-8.00;
Ion gun injection electric: 5000.00-5500.00;
Ion source temperature: 380.00-420.00.
3. assay method according to claim 1 is characterized in that: the said four unique nitrosamine internal standard compound standard items that grow tobacco are four deuterium generation-N-nitroso-nornicotines, four deuterium generation-N-nitroso-anabasines, four deuterium generation-N-nitroso-anatabines, four deuterium generation-4-(N-methyl-N-nitrosamine)-1-(3-pyridine radicals)-butanone.
4. according to claim 1 or 3 described assay methods, it is characterized in that: said four grow tobacco unique nitrosamine measured objects and the detected parameters of internal standard compound is following separately:
5. according to claim 1 or 3 described assay methods, it is characterized in that:
1. the preparation of standard reserving solution: accurately take by weighing the four unique nitrosamine standard items that grow tobacco respectively, mix, be mixed with concentration and respectively be the standard reserving solution of 0.8-1.2mg/mL with dissolve with methanol, constant volume;
2. the preparation of mark storing solution in: accurately take by weighing four respectively and grow tobacco the unique nitrosamine deuterium, mix, be mixed with concentration and respectively be the interior mark storing solution of 0.8-1.2mg/mL with dissolve with methanol, constant volume for the internal standard compound standard items;
6. assay method according to claim 5 is characterized in that:
The series standard solution of 7-8 variable concentrations gradient will be formulated as during detection after the standard reserving solution dilution; Concentration is in the 0-100ng/mL scope.
7. assay method according to claim 1 is characterized in that:
Cigarette mainstream flue gas sample pre-treatments: have the glass fibre filter disc of cigarette sample main flume to be positioned in the conical flask of 50mL ground band plug capture, add the 10-15mL extract, get the 0.8-1.2mL extract behind the mechanical oscillation 20-40min, filter back usefulness to be analyzed.
8. assay method according to claim 7 is characterized in that: filter said filter disc extraction back is to adopt 0.22 μ m water membrane filtration.
9. assay method according to claim 1 is characterized in that:
Utilize the high performance liquid chromatography-tandem mass appearance that series standard solution is detected respectively; Four the grow tobacco chromatographic peak area and the corresponding internal standard compound peak area ratios of unique nitrosamine that obtain are right; Draw respectively four grow tobacco unique nitrosamine typical curve and obtain regression equation; Under the same conditions sample is detected then; To be measured four chromatographic peak areas that grow tobacco unique nitrosamine that obtain are brought in the corresponding regression equation, can be obtained in the sample corresponding four content that grow tobacco unique nitrosamine.
10. assay method according to claim 1 is characterized in that:
Step By Condition to the suction of cigarette when said cigarette mainstream flue gas is collected is: the cigarette of choosing is propped up and glass fibre filter disc balance 48 hours under 22 ± 1 ℃, relative humidity 60 ± 2% conditions; Under this condition, use smoking machine that cigarette is aspirated then; Capture cigarette smoke with the glass fibre filter disc; Every mouthful was aspirated 2 seconds; Draw volume 35mL; Every mouthful of interval 58 seconds, every group of suction 4-5 props up cigarette.
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| CN108387654A (en) * | 2018-02-09 | 2018-08-10 | 深圳瀚星翔科技有限公司 | The detection method of NNN and NNK in a kind of atomization vapour of electronic cigarette release |
| CN111272915A (en) * | 2020-03-28 | 2020-06-12 | 中国烟草总公司郑州烟草研究院 | Method for detecting four nitrosoamino acids in buccal cigarette by liquid chromatography-tandem mass spectrometry |
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| CN103776944A (en) * | 2014-01-13 | 2014-05-07 | 红云红河烟草(集团)有限责任公司 | Method for detecting NNAL (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol) and NNA in cut tobacco and cigarette smoke |
| CN103776944B (en) * | 2014-01-13 | 2015-07-15 | 红云红河烟草(集团)有限责任公司 | Method for detecting NNAL (4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol) and NNA in cut tobacco and cigarette smoke |
| CN103837633A (en) * | 2014-03-16 | 2014-06-04 | 国家烟草质量监督检验中心 | Liquid chromatogram tandem mass spectrum measurement method for tobacco-specific nitrosamines in electronic cigarette liquid |
| CN104122357A (en) * | 2014-07-22 | 2014-10-29 | 云南中烟工业有限责任公司 | Method for detecting specific nitrosamine (TSNAs) in tobacco under condition of Canada depth suction |
| CN104237431B (en) * | 2014-08-15 | 2016-05-25 | 云南中烟工业有限责任公司 | The assay method of TSNAs in a kind of electronic cigarette smog |
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| CN104749310A (en) * | 2015-04-13 | 2015-07-01 | 中国烟草总公司四川省公司 | Method for measuring content of nitrosamine in cigarette filter |
| CN104749310B (en) * | 2015-04-13 | 2016-06-29 | 中国烟草总公司四川省公司 | A kind of measure the method for content of nitrosamines in cigarette filter |
| CN105938120A (en) * | 2016-07-18 | 2016-09-14 | 中国烟草总公司郑州烟草研究院 | Mass spectrum method for online detecting solanesol in cigarette environment smoke |
| CN106442753A (en) * | 2016-08-30 | 2017-02-22 | 中国烟草总公司四川省公司 | Method for measuring content of TSNAs (tobacco-specific nitrosamines) in mainstream smoke of cigarettes |
| CN108387654A (en) * | 2018-02-09 | 2018-08-10 | 深圳瀚星翔科技有限公司 | The detection method of NNN and NNK in a kind of atomization vapour of electronic cigarette release |
| CN111272915A (en) * | 2020-03-28 | 2020-06-12 | 中国烟草总公司郑州烟草研究院 | Method for detecting four nitrosoamino acids in buccal cigarette by liquid chromatography-tandem mass spectrometry |
| CN112816283A (en) * | 2020-12-30 | 2021-05-18 | 国家烟草质量监督检验中心 | Preparation method of tobacco specific N-nitrosamine standard substance |
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