CN106645441B - A method of evaluation raw tobacco material carbonyls burst size - Google Patents
A method of evaluation raw tobacco material carbonyls burst size Download PDFInfo
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- CN106645441B CN106645441B CN201610829665.2A CN201610829665A CN106645441B CN 106645441 B CN106645441 B CN 106645441B CN 201610829665 A CN201610829665 A CN 201610829665A CN 106645441 B CN106645441 B CN 106645441B
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- 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
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- G—PHYSICS
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- 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
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Abstract
The present invention relates to a kind of methods for evaluating raw tobacco material carbonyls burst size, belong to technical field of analytical chemistry.The present invention mainly comprises the steps of: sample dissociation；Product trapping；Product extraction；Product detection.The condition wherein cracked are as follows: using nitrogen oxygen atmosphere as carrier gas, be warming up to 600~1200 DEG C under 60~120 DEG C/s rate, then keep 2~10s.Nitrogen oxygen atmosphere is made of the oxygen and surplus nitrogen of volume fraction 9%, 16~17mL/s of flow rate of carrier gas, the condition can crack atmosphere when atmosphere simulation cigarette smoking with raw tobacco material, the carbonyl compound content generated when the burst size of raw tobacco material carbonyls and its manufactured cigarette smoking has preferable consistency, it is horizontal that its carbonyls burst size can be evaluated without cigarette manufacture, save the cost, while data support is provided to reduce carbonyl compound content technical research in cigarette.
Description
Technical field
The present invention relates to a kind of methods for evaluating raw tobacco material carbonyls burst size, belong to technique of analytical chemistry neck
Domain.
Background technique
Volatile carbonyl compound is a kind of important harmful substance in cigarette smoke, is had to human respiratory strong
Stimulation, such compound (formaldehyde, acetaldehyde, propionic aldehyde, methacrylaldehyde, propionic aldehyde, crotonaldehyde, 2- butanone and butyraldehyde) are put into
Hoffmann cigarette smoke nocuousness list.Wherein, formaldehyde, acetaldehyde and crotonaldehyde are arranged by international cancer research institution (IARC) respectively
For 1/2B, 3 class human carcinogens.Carbonyl compound content level in various raw tobacco materials is grasped, different tobacco leaves are rationally compounded
And various carbonyl compound content regulations are of great significance in cigarette.
Currently, detection tobacco product in content of volatile carbonyl compound mainly use spectrophotometry, paper chromatography,
The methods of high performance liquid chromatography and gas chromatography.A kind of method of common trapping carbonyls is in acid condition
2,4-dinitrophenylhydrazine (DNPH) is reacted with carbonyls generates stable hydrazone compounds, the hydrazone class chemical combination being subsequently formed
Object and then can determine the content of various carbonyls by UV detector in the wavelength detecting of 365-380nm.
Summary of the invention
The present invention provides a kind of methods for evaluating raw tobacco material carbonyls burst size, and its object is to tobacco leaf original
Expect that carbonyls burst size of the cigarette made of it in suction can be grasped without cigarette manufacture.
It is a kind of evaluate raw tobacco material carbonyls burst size method comprise the steps of: 1) sample dissociation；2) product
Trapping: sample material generates carbonyls by cracking, then traps to product；3) product extracts；4) product detection:
Extract liquor is separated and quantified with high performance liquid chromatography.
The method of the evaluation raw tobacco material carbonyls burst size, further includes Preparatory work of experiment before sample dissociation,
The Preparatory work of experiment mainly includes raw tobacco material pretreatment, 2,4 dinitrophenyl hydrazine recrystallization, 2,4 dinitrophenyl hydrazine saturation
Solution is prepared, the preparation of 2,4 dinitrophenyl hydrazine filter disc, extract liquor is prepared and standard working solution is prepared.
Raw tobacco material pretreatment for according to GB/T 16447-2004 " tobacco and tobacco product adjust and test it is big
Compression ring border " requirement, tobacco sample is in 22 ± 2 DEG C of temperature and the lower balance 48h of relative humidity 60 ± 5%.
The 2,4-dinitrophenylhydrazine recrystallization heats to take 50g 2,4-dinitrophenylhydrazine to be added in 800mL acetonitrile
It is back to 2,4-dinitrophenylhydrazine to be completely dissolved, cooled to room temperature, stand, filter, acetonitrile washing, vacuum drying.
The 2,4 dinitrophenyl hydrazine saturated solution be formulated as weighing the 2,4 dinitrophenyl hydrazine solid of 7.5g recrystallization in
In brown boiling flask, it is separately added into 500mL acetonitrile and 1mL70% perchloric acid, heating and ultrasound extremely dissolution, cold under the conditions of 60 DEG C
But to room temperature to obtain the final product.
The 2,4-dinitrophenylhydrazine filter disc is prepared as being that 44mm glass fiber filter is stacked by two diameters, is uniformly added into
2mL 2,4-dinitrophenylhydrazine saturated solution is placed in be protected from light in vacuum desiccator and dry.Before pipe tobacco cracking, in the filter dried
It is stand-by that 1mL 2,4 dinitrophenyl hydrazine saturated solution is added on piece.
The extract liquor is formulated as pyridine/acetonitrile solution that dose volume score is 2%.
The standard working solution is to contain following 8 kinds of phenylhydrazone compounds: formaldehyde -2,4- dinitrophenylhydrazone, acetaldehyde -2,4-
Dinitrophenylhydrazone, acetone -2,4- dinitrophenylhydrazone, methacrylaldehyde -2,4- dinitrophenylhydrazone, propionic aldehyde -2,4- dinitrophenylhydrazone, crotons
Aldehyde -2,4- dinitrophenylhydrazone, 2- butanone-2, the series standard working solution of 4- dinitrophenylhydrazone and butyraldehyde -2,4- dinitrophenylhydrazone.
The preparation method of the standard working solution is as follows: 1) preparing standard reserving solution: weighing formaldehyde -2,4- dinitro respectively
Base phenylhydrazone 50mg, acetaldehyde -2,4- dinitrophenylhydrazone 100mg, acetone -2,4- dinitrophenylhydrazone 50mg, methacrylaldehyde -2,4- dinitro
Phenylhydrazone 40mg, propionic aldehyde -2,4- dinitrophenylhydrazone 30mg, crotonaldehyde -2,4- dinitrophenylhydrazone 30mg, 2- butanone-2,4- dinitro
Phenylhydrazone 30mg and butyraldehyde -2,4- dinitrophenylhydrazone 30mg is added in 50mL beaker, after addition 30mL acetonitrile is completely dissolved, is transferred to
In the volumetric flask of 100mL, it is settled to scale with acetonitrile, obtains standard reserving solution；2) it prepares primary standard liquid: pipetting the storage of 10mL standard
Standby liquid is settled to scale with acetonitrile, obtains primary standard liquid into 50mL volumetric flask；3) secondary standard liquid is prepared: accurate respectively to move
Take 0.1,0.2,0.5,1,5,10,25mL primary standard liquid into 50mL volumetric flask, be settled to scale with acetonitrile, obtain 7 second levels
Titer.Primary standard liquid and 7 secondary standard liquid collectively form series standard working solution.
In the method for the evaluation raw tobacco material carbonyls burst size, sample dissociation is to repeat to crack two parts of samples
Product, cracking condition are as follows: using nitrogen oxygen atmosphere as carrier gas, 600~1200 DEG C are warming up under 60~120 DEG C/s rate, then keep 2~
10s.Nitrogen oxygen atmosphere is made of the oxygen and surplus nitrogen of volume fraction 9%, 16~17mL/s of flow rate of carrier gas.
The condition can crack atmosphere when atmosphere simulation cigarette smoking with raw tobacco material, and raw tobacco material carbonyls is released
High-volume with its made of cigarette smoking when the carbonyls measurer that generates have preferable consistency, can be cracked with raw tobacco material
The carbonyl compound content of generation evaluates the carbonyl compound content generated when its manufactured cigarette smoking.
In the method for the evaluation raw tobacco material carbonyls burst size, extract liquor used in sample extraction is volume
Pyridine/acetonitrile solution that score is 2%, the volume of extract liquor used are 100mL.
In the method for the evaluation raw tobacco material carbonyls burst size, high-efficient liquid phase chromatogram condition in product detection
Are as follows: detector: diode array detector；Detection wavelength: 365nm；Sample volume: 10 μ L；Column oven temperature: 30 DEG C；Chromatographic column:Explosive E2 250mm×4.6mm5 μm of chromatographic column filler granularity；Mobile phase: A is deionized water, B
For acetonitrile, flow velocity: 1mL/min；Elution requirement: 0~20min, 50%A-50%A, 20~25min, 50%A-40%A, 25~
30min, 40%A-40%A, 30~35min, 40%A-20%A, 35~40min, 20%A-10%A, 40~41min, 10%
A-50%A, 41~45min, 50%A-50%A.
Compared with the prior art, the advantages of the present invention are as follows:
Raw tobacco material cracking atmosphere of the invention is similar to condition when cigarette smoking, can be cracked and be produced by raw tobacco material
Raw carbonyl compound content evaluates carbonyl compound content when its manufactured cigarette smoking, to screening raw tobacco material and tobacco leaf
Raw material compounding, which is of great significance and reduces technical research for carbonyls burst size in cigarette, provides data reference and skill
Art is supported, the carbonyl compound content in raw tobacco material can be evaluated without cigarette, has saved cost.
The present invention makes filter disc using oversaturated 2,4-dinitrophenylhydrazine solution, the volatilization that can be generated to raw tobacco material
Property carbonyls more effectively trapped, while using recrystallization 2,4-dinitrophenylhydrazine, 2,4- bis- can be effectively reduced
The content of carbonyl compound impurities, reduces background interference in nitrophenyl hydrazine solid, keeps measurement result more accurate.
Present invention extract liquor used in product extraction process is pyridine/acetonitrile mixture that volume fraction is 2%, used
Extract liquor volume be 100mL, make to reach balance between extraction efficiency and consumption of organic solvent, not only improve extraction efficiency, but also
Consumption of organic solvent is saved.
Detailed description of the invention
Fig. 1 is the high-efficient liquid phase chromatogram for detecting sample and standard working solution, wherein 1 is formaldehyde -2,4- dinitrophenylhydrazone, 2
It is acetone -2,4- dinitrophenylhydrazone for acetaldehyde -2,4- dinitrophenylhydrazone, 3,4 be methacrylaldehyde -2,4- dinitrophenylhydrazone, 5 is third
Aldehyde -2,4- dinitrophenylhydrazone, 6 be crotonaldehyde -2,4- dinitrophenylhydrazone, 7 be 2- butanone-2,4- dinitrophenylhydrazone and 8 is butyraldehyde -
The chromatographic peak of 2,4- dinitrophenylhydrazone.
Specific embodiment
It is more prominent with advantage in order to keep the solution of the present invention more specific, it is done furtherly below with reference to specific example
It is bright, but it is not intended to limit the present invention.
Instrument and reagent
High performance liquid chromatograph, acetonitrile (chromatographically pure), balance, formaldehyde, acetaldehyde, acetone, methacrylaldehyde, propionic aldehyde, crotonaldehyde, 2-
Butanone, butyraldehyde 2,4- dinitrophenylhydrazone standard items；2,4 dinitrophenyl hydrazine, pyridine, perchloric acid (analysis is pure).
Embodiment
Include Preparatory work of experiment, sample dissociation, product trapping, product extraction and product detection in the present embodiment.
1, Preparatory work of experiment
Preparatory work of experiment in the present embodiment mainly includes raw tobacco material pretreatment, 2,4 dinitrophenyl hydrazine recrystallization, 2,4-
Dinitrophenylhydrazine saturated solution is prepared, the preparation of 2,4 dinitrophenyl hydrazine filter disc, extract liquor is prepared and standard working solution is prepared
(a) raw tobacco material pre-processes: according to " the atmosphere of tobacco and tobacco product adjusting and test of GB/T 16447-2004
Environment " requirement, raw tobacco material is in 22 ± 2 DEG C of temperature and the lower balance 48h of relative humidity 60 ± 5%.
(b) 2,4-dinitrophenylhydrazine recrystallizes: taking 50g 2,4-dinitrophenylhydrazine to be added in 800ml acetonitrile, heats back
It flow to 2,4-dinitrophenylhydrazine to be completely dissolved, is transferred in beaker sealing and is protected from light natural cooling, stand overnight, crystallization filtered,
Acetonitrile washing, after vacuum drying to obtain the final product.
(c) 2,4 dinitrophenyl hydrazine saturated solution is prepared: weighing the purified 2,4 dinitrophenyl hydrazine solid of 7.5g in palm fibre
In the flat bottle of color, it is separately added into 500mL acetonitrile and 1mL70% perchloric acid, heating and ultrasound extremely dissolution, are cooled under the conditions of 60 DEG C
Room temperature obtains 2,4 dinitrophenyl hydrazine saturated solution；
(d) prepared by 2,4 dinitrophenyl hydrazine filter disc: 2,4 dinitrophenyl hydrazine filter disc is prepared as two diameters being 44mm glass
Glass fiber filter disc stacks, and is uniformly added into 2mL2, and 4- dinitrophenylhydrazine saturated solution is placed in be protected from light in vacuum desiccator and dry.
Before pipe tobacco cracking, 1mL2 is added on the filter disc dried, 4- dinitrophenylhydrazine saturated solution is stand-by.
(e) extraction solution is prepared: pyridine/acetonitrile solution of dose volume score 2%.
(f) preparation of standard working solution:
1) it prepares standard reserving solution: weighing formaldehyde -2,4- dinitrophenylhydrazone 50mg, acetaldehyde -2,4- dinitrophenylhydrazone respectively
100mg, acetone -2,4- dinitrophenylhydrazone 50mg, methacrylaldehyde -2,4- dinitrophenylhydrazone 40mg, propionic aldehyde -2,4- dinitrophenylhydrazone
30mg, crotonaldehyde -2,4- dinitrophenylhydrazone 30mg, 2- butanone-2,4- dinitrophenylhydrazone 30mg and butyraldehyde -2,4- dinitrophenylhydrazone
30mg is added in 50mL beaker, after addition 30mL acetonitrile is completely dissolved, is transferred in the volumetric flask of 100mL, is settled to acetonitrile
Scale obtains standard reserving solution；2) it prepares primary standard liquid: pipetting 10mL standard reserving solution into 50mL volumetric flask, it is fixed with acetonitrile
Hold to scale, obtains primary standard liquid；3) it prepares secondary standard liquid: accurately pipetting 0.1,0.2,0.5,1,5,10,25mL mono- respectively
Grade standard liquid is settled to scale into 50mL volumetric flask, with acetonitrile, obtains 7 secondary standard liquid.Primary standard liquid and 7 second level marks
Quasi- liquid collectively forms standard working solution.
2, sample dissociation
Repeat to crack 2 parts of tobacco sample, every part of sample quality is 50~200mg, cracking condition are as follows: temperature program: Carrier gas: 9%O2(volume fraction, remaining is N2), 16.7mL/s.
3, product traps
Cracking finishes, and removes trap, places 3min, make carbonyls in flue gas sufficiently with 2,4- dinitro filter disc
Sufficiently effect.
4, product extracts
Filter disc is taken out, is transferred in the conical flask for filling 100mL extraction solution, 160~200 revs/min, mechanical oscillation
15min stands 2min, and extract liquor crosses 0.45 μm of organic phase filter membrane, analyzes into HPLC.
5, product detection
It to standard working solution and is trapped product with high performance liquid chromatography and analyzed, high-efficient liquid phase chromatogram condition are as follows:
Detector: diode array detector；
Detection wavelength: 365nm；
Sample volume: 10 μ L；
Column oven temperature: 30 DEG C；
Chromatographic column:Explosive E2 250mm×4.6mm5 μm of chromatographic column filler granularity；
Mobile phase: A: deionized water, B: acetonitrile, flow velocity: 1mL/min；Gradient see the table below 1
1 gradient table of table
| Time, min | Mobile phase A, % | Mobile phase B, % |
| 0 | 50 | 50 |
| 20 | 50 | 50 |
| 25 | 40 | 60 |
| 30 | 40 | 60 |
| 35 | 20 | 80 |
| 40 | 10 | 90 |
| 41 | 50 | 50 |
| 45 | 50 | 50 |
Experimental result and analysis
Each corresponding concentration of carbonyl compound peak area that analysis obtains is made into regression analysis, obtains the line of concentration and peak area
Property equation, the range of linearity and related coefficient, see the table below 2；
The linear equation and the range of linearity of each carbonyl compound of table 2
| Serial number | Detect ingredient | Linear equation | The range of linearity | Related coefficient |
| 1 | Formaldehyde | Y=342.02x-0.52 | 0.03~14.96 | 0.9999 |
| 2 | Acetaldehyde | Y=259.64x-0.30 | 0.08~39.09 | 1.0000 |
| 3 | Acetone | Y=196.04x+0.12 | 0.05~27.25 | 0.9999 |
| 4 | Methacrylaldehyde | Y=251.20x+0.24 | 0.04~19.38 | 0.9999 |
| 5 | Propionic aldehyde | Y=202.46x+0.22 | 0.03~16.90 | 1.0000 |
| 6 | Crotonaldehyde | Y=181.35x+0.63 | 0.04~19.09 | 1.0000 |
| 7 | 2- butanone | Y=150.92x+0.41 | 0.03~17.26 | 1.0000 |
| 8 | Butyraldehyde | Y=165.44x+0.24 | 0.03~17.20 | 0.9999 |
From table 2 as can be seen that eight kinds of carbonyl compound linear relationship wider range, related coefficient between 0.9999~1.0,
Linear equation can be used to the carbonyl compound content in accurate evaluation raw tobacco material.
3 raw tobacco material thermal cracking of table and cigarette smoking carbonyls burst size Comparative result
Cigarette is made in control tobacco sample, compares tobacco sample thermal cracking and cigarette smoking result (being shown in Table 3), 8 kinds of carbonyls
Based compound thermal cracking and cigarette smoking result ratio range are 95%~107%, it can be seen that, raw tobacco material carbonyls
Burst size and the carbonyl compound content that generates of when its manufactured cigarette smoking there is preferable consistency.
4 different sources of table and position raw tobacco material carbonyls burst size (mg/g)
From table 4, it can be seen that various carbonyls in the raw tobacco material of different sources and the raw tobacco material of different parts
Content it is different, the content of various carbonyls is different in tobacco material, wherein acetaldehyde highest, be 4.98~
7.58mg/g, content of formaldehyde take second place, and are 2.47~4.96mg/g, and acetone and level of acrolein are suitable, respectively 1.36~
2.16mg/g and 1.06~2.36mg/g, the content of propionic aldehyde are 0.35~0.81mg/g, and crotonaldehyde, 2- butanone and butyraldehyde are compared
Yu Qiansi kind, content is less, is followed successively by 0.21~0.55mg/g, 0.12~0.41mg/g and 0.11~0.36mg/g.These numbers
It is of great significance according to screening raw tobacco material and raw tobacco material compounding, while also being reduced for carbonyls burst size in cigarette
Technical research provides data reference and technical support.
Claims (3)
1. a kind of method for evaluating raw tobacco material carbonyls burst size, it is characterised in that: comprise the steps of: that sample is split
Solution；Product trapping: sample material generates carbonyls by cracking, then traps to product；Product extraction；Product inspection
It surveys: extract liquor being separated and quantified with high performance liquid chromatography；
It further include Preparatory work of experiment before sample dissociation, the Preparatory work of experiment mainly includes raw tobacco material pretreatment, 2,4- dinitro
Phenylhydrazine recrystallization, 2,4 dinitrophenyl hydrazine solution are prepared, the preparation of 2,4 dinitrophenyl hydrazine filter disc, extract liquor is prepared and standard work
Liquid is prepared；The 2,4 dinitrophenyl hydrazine solution is supersaturated solution；
The 2,4-dinitrophenylhydrazine recrystallization is heated to reflux to take 50g 2,4-dinitrophenylhydrazine to be added in 800mL acetonitrile
It is completely dissolved, cooled to room temperature, stands to 2,4-dinitrophenylhydrazine, filter, acetonitrile washing, vacuum drying；
The 2,4 dinitrophenyl hydrazine supersaturated solution is formulated as weighing the 2,4 dinitrophenyl hydrazine solid of 7.5g recrystallization in palm fibre
In color boiling flask, it is separately added into 500mL acetonitrile and 1mL70% perchloric acid, heating and ultrasound extremely dissolution, cooling under the conditions of 60 DEG C
To room temperature to obtain the final product；
The extract liquor is formulated as pyridine/acetonitrile solution that dose volume score is 2%；
The sample dissociation is to repeat to crack two parts of samples, cracking condition are as follows: using nitrogen oxygen atmosphere as carrier gas, from 35 DEG C 90
DEG C/s rate under be warming up to 900 DEG C, then keep 6s, nitrogen oxygen atmosphere is made of the oxygen and surplus nitrogen of volume fraction 9%, is carried
Gas velocity 16.7mL/s；
High-efficient liquid phase chromatogram condition in the product detection are as follows: detector: diode array detector；Detection wavelength: 365
nm；Sample volume: 10 μ L；Column oven temperature: 30 DEG C；Chromatographic column: Acclaim® Explosive E2 250mm×4.6mm 120
, 5 μm of chromatographic column filler granularity；Mobile phase: A is deionized water, and B is acetonitrile, flow velocity: 1mL/min, elution requirement: 0 ~ 20min,
50%A-50%A, 20 ~ 25min, 50%A-40%A, 25 ~ 30min, 40%A-40%A, 30 ~ 35min, 40%A-20%A, 35 ~ 40min,
20%A-10%A, 40 ~ 41min, 10%A-50%A, 41 ~ 45min, 50%A-50%A.
2. the method for evaluation raw tobacco material carbonyls burst size according to claim 1, it is characterised in that: described
Standard working solution contains following 8 kinds of phenylhydrazone compounds: formaldehyde -2,4- dinitrophenylhydrazone, acetaldehyde -2,4- dinitrophenylhydrazone, acetone -
2,4- dinitrophenylhydrazone, methacrylaldehyde -2,4- dinitrophenylhydrazone, propionic aldehyde -2,4- dinitrophenylhydrazone, crotonaldehyde -2,4- dinitrobenzene
Hydrazone, 2- butanone-2, the series standard working solution of 4- dinitrophenylhydrazone and butyraldehyde -2,4- dinitrophenylhydrazone.
3. the method for evaluation raw tobacco material carbonyls burst size according to claim 1, it is characterised in that: described
Product extraction extract liquor used is pyridine/acetonitrile solution that volume fraction is 2%, and the volume of extract liquor used is 100 mL.
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