CN105891383A - GC-MS/MS method for synchronously detecting specific nitrosamine and volatile N-nitrosamine in tobacco and tobacco products - Google Patents
GC-MS/MS method for synchronously detecting specific nitrosamine and volatile N-nitrosamine in tobacco and tobacco products Download PDFInfo
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- CN105891383A CN105891383A CN201610386484.7A CN201610386484A CN105891383A CN 105891383 A CN105891383 A CN 105891383A CN 201610386484 A CN201610386484 A CN 201610386484A CN 105891383 A CN105891383 A CN 105891383A
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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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Abstract
The invention discloses a GC-MS/MS method for synchronously detecting specific nitrosamine and volatile N-nitrosamine in tobacco and tobacco products. Four kinds of nitrosamine of NNN, NAT, NAB and NNK and eight kinds of volatile N-nitrosamine of NDMA, NPYR, NPIP, NMOR, NRMA, NDEA, NDPA and NDBA are specially included. The method is characterized in that a sodium hydroxide solution is added into a tobacco sample, dichloromethane is added after uniform mixing carried out in a handholding oscillation mode, oscillation is carried out through an oscillator, anhydrous magnesium sulfate is added, the mixture is added into the oscillator to be oscillated after handholding oscillation, then, centrifugation and concentration are carried out, and finally, GC-MS/MS analysis is carried out. The method has the advantages that sample pretreatment is easy and convenient, the specific nitrosamine and volatile N-nitrosamine in the tobacco can be analyzed synchronously, batch treatment can be carried out, the measuring speed is high, the adding standard recovery rate is good, the degree of precision is high, and the technical support can be provided for accurately assessing the safety risks of the specific nitrosamine and volatile N-nitrosamine in the tobacco and tobacco products.
Description
Technical field
The invention belongs to tobacco-specific nitrosamine and volatile n-nitrosamine detection technique field in tobacco and tobacco product,
Specifically use gas chromatography tandem mass spectrometry method (GC-MS/MS) to analyze 4 kinds of peculiar Asias of tobacco in tobacco and tobacco product simultaneously
Nitramine and 8 kinds of volatile n-nitrosamines, utilize the method can 12 kinds of nitrosamine in fast quantitative analysis tobacco and tobacco product
Content.
Background technology
Tobacco-specific nitrosamine (TSNAs) is a topmost class carcinogen in tobacco and tobacco product, and in TSNAs
4 kinds paid close attention to most are respectively N-nitrosonornicotine (NNN), N-nitrosoanatabine (NAT), N-nitroso anabasine
(NAB), 4-methyl nitroso pyridine radicals butanone (NNK)；Volatile n-nitrosamine (VNAs) is the cause that a class receives significant attention
Cancer class material, before 1970, researcher just has been found that in tobacco containing volatile n-nitrosamine, mainly includes that N-is sub-
Nitro dimethyl amine (NDMA), N-nitrosopyrolidine (NPYR), N-nitroso-piperidine (NPIP), N-nitrosomorpholine
(NMOR), N-nitroso Methylethyl amine (NEMA), N-Nitrosodiethylamine (NDEA), N-nitroso di-n-propylamine (NDPA) and N-
8 kinds of volatile n-nitrosamines such as nitroso dibutyl amine (NDBA), wherein NDMA is 2A level carcinogenic substance, and NPYP, NMOR, NPIP are 2B
Level carcinogenic substance.
The matrix of tobacco and tobacco product is more complicated, needs to use corresponding pre-treating method in the detection of nitrosamine
Decontamination substrate.The pretreatment technologies such as SPE, liquid-liquid extraction, SPME can reduce matrix effect, but the most also
Add sample analysis time and limit sample throughput, so far, about measuring tobacco in tobacco and tobacco product simultaneously
The report of unique nitrosamine and volatile n-nitrosamine is little, it is necessary to study further.
Summary of the invention
The one that the purpose of the present invention is based on above-mentioned prior art situation and provides specially detects tobacco and cigarette simultaneously
The GC-MS/MS method of tobacco-specific nitrosamine and volatile n-nitrosamine in straw-made articles, it is achieved that to tobacco-specific nitrosamine and
Analyze while volatile n-nitrosamine, simplify pre-treatment operation, tobacco and tobacco product can be carried out batch quickly inspection
Survey.
It is an object of the invention to be achieved through the following technical solutions:
Tobacco-specific nitrosamine and the GC-MS/MS method of volatile n-nitrosamine in a kind of detection tobacco simultaneously and tobacco product,
Specifically include four to grow tobacco unique nitrosamine: NNN, NAT, NAB, NNK, and eight kinds of volatile n-nitrosamine: NDMA, NPYR,
NPIP, NMOR, NEMA, NDEA, NDPA, NDBA, be addition sodium hydroxide solution in tobacco sample, and hand-held vibration mixes
Rear addition dichloromethane, oscillator vibrates, and adds anhydrous magnesium sulfate, and hand-held vibration is placed in oscillator vibration, be then centrifuged for,
Concentrate, finally analyze with GC-MS/MS, specifically comprise the following steps that
A, the preparation of inner mark solution: preparation internal standard concentration is NDMA-D6, NMEA-D3, NDEA-D10, NDPA-of 1 g/ml
The mixing internal standard dichloromethane working solution of D14, NMOR-D8, NPYR-D8, NPIP-D8, NDBA-D8 and NNK-D4, NAT-D4,
The mixing internal standard dichloromethane solution of NAB-D4 and NNK-D4；
B, the preparation of mixed standard solution: with dichloromethane as solvent, prepare tobacco-specific nitrosamine series standard solution, specifically
Concentration is: 5 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml, 500 ng/ml, 1000 ng/ml,
5000 ng/ml；It is separately added into tobacco-specific nitrosamine inner mark solution, makes the concentration of each internal standard compound be 400 ng/ml；With two
Chloromethanes is solvent, prepares volatile n-nitrosamine series standard solution,Concrete concentration is: 0.2 ng/mL, 0.5 ng/mL, 1
Ng/mL, 2 ng/mL, 5 ng/mL, 10 ng/mL, 20 ng/mL, 50 ng/ml and 100 ng/ml, be separately added into volatility N-
Nitrosamine inner mark solution, makes the concentration of each internal standard compound be 20 ng/ml.
C, tobacco sample prepare: tobacco and tobacco product are stored refrigerated with standby after grinding；
D, extraction: the 4 g samples accurately weighed are put in 50 mL centrifuge tubes that to add the NaOH that 4 ml concentration are 10% molten
Liquid is to improve the extraction effect of object, and hand-held vibration 15 s mixes, and is subsequently adding 30 ml dichloromethane interior with corresponding
(addition of tobacco-specific nitrosamine inner mark solution is 200 l to mark solution, the addition 20 of volatile n-nitrosamine inner mark solution
L), centrifuge tube being placed in oscillator and vibrates 20 min, add 4.5 g anhydrous magnesium sulfates after vibration, hand-held vibration 15 s is placed on
Vibrate in oscillator 2 min；
E, centrifugal: sample is centrifuged 5 min with 11000 r/min on supercentrifuge；
F, concentration: take supernatant nitrogen at 43 DEG C ± 2 DEG C after Li Xin and blow and be concentrated into about 2 ml, transfer them to 2 mL chromatograms little
In Ping, in case detection；
G, GC-MS/MS measure, GC-MS/MS Instrument working parameter such as table 1:
Table 1 GC-MS/MS Instrument working parameter
Object and interior target retention time and mass spectrometry parameters such as table 2:
Table 2 object and interior target retention time and mass spectrometry parameters
| Retention time (min) | Quota ion pair | Qualitative ion pair | |
| NDMA | 8.758 | 92→75 | 92→58 |
| NDMA-D6 | 8.715 | 98→81 | 98→46 |
| NMEA | 10.528 | 106→89 | 106→61 |
| NMEA-D3 | 10.504 | 109→92 | 109→64 |
| NDEA | 11.908 | 120→103 | 120→75 |
| NDEA-D4 | 11.822 | 130→113.1 | 130→81 |
| NMBA | 14.150 | 134→117 | 134→61 |
| NDPA | 15.013 | 148→131.1 | 148→89 |
| NDPA-D10 | 14.891 | 162→145 | 162→50 |
| NMOR | 15.173 | 134→117 | 134→87 |
| NMOR-D8 | 15.125 | 142→125 | 142→95 |
| NPYR | 15.390 | 118→101 | 118→55 |
| NPYR-D10 | 15.322 | 126→109 | 126→62 |
| NPIP | 15.960 | 132→115 | 132→69 |
| NPIP-D10 | 15.893 | 142→125 | 142→78 |
| NDBA | 18.014 | 176→159 | 176→103 |
| NDBA-D18 | 17.858 | 194→177 | 194→113 |
| NNN | 26.645 | 178→148 | 178→120.5 |
| NNN-D4 | 26.599 | 182→109 | 182→152 |
| NAT | 27.266 | 190→160.1 | 190→106 |
| NAT-D4 | 27.211 | 194→164 | 194→110 |
| NAB | 27.978 | 192→162 | 192→133.1 |
| NAB-D4 | 27.923 | 196→166 | 196→136 |
| NNK | 32.110 | 208→122.1 | 208→148 |
| NNK-D4 | 32.026 | 212→126 | 212→152 |
H, result calculate.
It is an advantage of the current invention that: sample pre-treatments simplicity, tobacco-specific nitrosamine and volatile n-nitrosamine can be same
Time analyzed, can batch processing, and the method finding speed is fast, recovery of standard addition good, precision is high, can be accurate evaluation tobacco
And the security risk of tobacco-specific nitrosamine and volatile n-nitrosamine provides technical support in tobacco product.
Detailed description of the invention
The present invention combines example and is further described, but is not limiting as the present invention.
Example 1
According to step (b) preparation series standard solution in foregoing invention content, it is measured with GC-MS/MS the most again, obtains line
Property equation and coefficient correlation.It is 3 and 10 calculating detection limit and quantitative limit with signal to noise ratio.Result shows method good linearity, detection limit
Low (the results are shown in Table 3)
Table 3 tobacco-specific nitrosamine and the calibration curve of volatile n-nitrosamine, detection limit and quantitative limit
Example 2
According to step d, e and f in foregoing invention content, before sample treatment, add quantitative standard solution, result such as table 4 institute
Showing, the in a few days rate of recovery of all objects is between 88.5%-117%, and the rate of recovery is between 89.2%-107% in the daytime.In a few days accurate
Degree and day to day precision are respectively 4.3%-13.5% and 6.2%-15.1%, and precision is good.
Example 3
According to the tobacco-specific nitrosamine in 7 kinds of samples of step measurements in foregoing invention content and volatile n-nitrosamine content,
The results are shown in Table 5.
。
Claims (3)
1. one kind is detected tobacco-specific nitrosamine and the GC-MS/MS side of volatile n-nitrosamine in tobacco and tobacco product simultaneously
Method, specifically includes four and grows tobacco unique nitrosamine: NNN, NAT, NAB, NNK, and eight kinds of volatile n-nitrosamine: NDMA, NPYR,
NPIP、NMOR、NEMA、NDEA、NDPA、NDBA；It is characterized in that: in tobacco sample, add sodium hydroxide solution, hand-held shake
Swinging addition dichloromethane after mixing, oscillator vibrates, and adds anhydrous magnesium sulfate, and hand-held vibration is placed in oscillator shakes
Swing, be then centrifuged for, concentrate, finally analyze with GC-MS/MS.
GC-MS/MS method the most according to claim 1, it is characterised in that: specifically comprising the following steps that of the method
A, the preparation of inner mark solution: preparation internal standard concentration is NDMA-D6, NMEA-D3, NDEA-D10, NDPA-of 1 g/ml
The mixing internal standard dichloromethane solution of D14, NMOR-D8, NPYR-D8, NPIP-D8, NDBA-D8 and NNK-D4, NAT-D4, NAB-
The mixing internal standard dichloromethane solution of D4 and NNK-D4；
B, the preparation of mixed standard solution: with dichloromethane as solvent, prepare tobacco-specific nitrosamine series standard solution, and
It is separately added into tobacco-specific nitrosamine inner mark solution, makes the concentration of each internal standard compound be 400 ng/ml；It is molten with dichloromethane
Agent, prepares volatile n-nitrosamine series standard solution,And it is separately added into volatile n-nitrosamine inner mark solution, make each internal standard compound
Concentration be 20 ng/ml；
C, tobacco sample prepare: tobacco and tobacco product are stored refrigerated with standby after grinding；
D, extraction: the 4 g samples accurately weighed are put in 50 mL centrifuge tubes that to add the NaOH that 4 ml concentration are 10% molten
Liquid, hand-held vibration 15 s mixes, and is subsequently adding 30 ml dichloromethane and corresponding inner mark solution, is placed in by centrifuge tube and shakes
Swinging device to vibrate 20 min, add 4.5 g anhydrous magnesium sulfates after vibration, hand-held vibration 15 s is placed in oscillator 2 min that vibrate；
E, centrifugal: sample is centrifuged 5 min with 11000 r/min on supercentrifuge；
F, concentration: take supernatant nitrogen at 43 DEG C ± 2 DEG C after Li Xin and blow and be concentrated into about 2 ml, transfer them to 2 mL chromatograms little
In Ping, in case detection；
G, GC-MS/MS measure, GC-MS/MS Instrument working parameter such as following table:
H, result calculate.
GC-MS/MS method the most according to claim 1, it is characterised in that: tobacco-specific nitrosamine series standard solution
Concrete concentration is: 5 ng/ml, 10 ng/ml, 50 ng/ml, 100 ng/ml, 200 ng/ml, 500 ng/ml, 1000 ng/
ml、5000 ng/ml；The concrete concentration of volatile n-nitrosamine series standard solution is: 0.2 ng/mL, 0.5 ng/mL, 1
Ng/mL, 2 ng/mL, 5 ng/mL, 10 ng/mL, 20 ng/mL, 50 ng/ml and 100 ng/ml.
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Cited By (6)
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| CN106324157A (en) * | 2016-10-27 | 2017-01-11 | 安徽易康达光电科技有限公司 | Method for quickly detecting volatile N-nitrosamine in cosmetics |
| CN108760917A (en) * | 2018-05-24 | 2018-11-06 | 中国烟草总公司郑州烟草研究院 | One grow tobacco and the processing method of tobacco product, tobacco and tobacco product in N- nitrosodiethanolamine contents assay method |
| CN108956802A (en) * | 2018-05-24 | 2018-12-07 | 中国烟草总公司郑州烟草研究院 | The detection method of Multiple components in the processing method of a kind of tobacco and tobacco product, tobacco and tobacco product |
| CN114720591A (en) * | 2022-03-22 | 2022-07-08 | 北京安胜瑞力科技有限公司 | Quantitative analysis method for DMAC (dimethylacetamide) in industrial wastewater |
| CN115078619A (en) * | 2022-06-10 | 2022-09-20 | 重庆医科大学 | GC-MS/MS method for simultaneously detecting 9N-nitrosamine impurities in valsartan preparation |
| CN116735760A (en) * | 2023-08-16 | 2023-09-12 | 山东道可化学有限公司 | Method for detecting N, N-dipropyl nitrosamine in trifluralin |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106324157A (en) * | 2016-10-27 | 2017-01-11 | 安徽易康达光电科技有限公司 | Method for quickly detecting volatile N-nitrosamine in cosmetics |
| CN108760917A (en) * | 2018-05-24 | 2018-11-06 | 中国烟草总公司郑州烟草研究院 | One grow tobacco and the processing method of tobacco product, tobacco and tobacco product in N- nitrosodiethanolamine contents assay method |
| CN108956802A (en) * | 2018-05-24 | 2018-12-07 | 中国烟草总公司郑州烟草研究院 | The detection method of Multiple components in the processing method of a kind of tobacco and tobacco product, tobacco and tobacco product |
| CN108760917B (en) * | 2018-05-24 | 2020-11-24 | 中国烟草总公司郑州烟草研究院 | Tobacco and tobacco product treatment method and method for measuring content of N-nitrosodiethanolamine in tobacco and tobacco product |
| CN108956802B (en) * | 2018-05-24 | 2021-06-18 | 中国烟草总公司郑州烟草研究院 | Processing method of tobacco and tobacco products and detection method of various components in tobacco and tobacco products |
| CN114720591A (en) * | 2022-03-22 | 2022-07-08 | 北京安胜瑞力科技有限公司 | Quantitative analysis method for DMAC (dimethylacetamide) in industrial wastewater |
| CN114720591B (en) * | 2022-03-22 | 2022-12-30 | 北京安胜瑞力科技有限公司 | Quantitative analysis method for DMAC (dimethylacetamide) in industrial wastewater |
| CN115078619A (en) * | 2022-06-10 | 2022-09-20 | 重庆医科大学 | GC-MS/MS method for simultaneously detecting 9N-nitrosamine impurities in valsartan preparation |
| CN116735760A (en) * | 2023-08-16 | 2023-09-12 | 山东道可化学有限公司 | Method for detecting N, N-dipropyl nitrosamine in trifluralin |
| CN116735760B (en) * | 2023-08-16 | 2024-02-09 | 山东道可化学有限公司 | Method for detecting N, N-dipropyl nitrosamine in trifluralin |
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Application publication date: 20160824 |