CN103197020B - GC (Gas Chromatography)/MS (Mass Spectrometry) measuring method of humectants in cigarette cut tobacco - Google Patents
GC (Gas Chromatography)/MS (Mass Spectrometry) measuring method of humectants in cigarette cut tobacco Download PDFInfo
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- CN103197020B CN103197020B CN201310106355.4A CN201310106355A CN103197020B CN 103197020 B CN103197020 B CN 103197020B CN 201310106355 A CN201310106355 A CN 201310106355A CN 103197020 B CN103197020 B CN 103197020B
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Abstract
The invention discloses a GC (Gas Chromatography)/MS (Mass Spectrometry) measuring method of humectants in cigarette cut tobacco, wherein the humectants comprise 1,2-propylene glycol, glycerin and triglycol. The GC/MS measuring method is characterized by comprising the following steps of: drying, crushing and sieving a cut tobacco sample; respectively treating the extract liquor through a dispersive solid-phase extraction (SPE) centrifuge tube and purifying the extract liquor through a filter membrane, quantitatively detecting the contents of the three humectants in the cut tobacco through a gas chromatography and mass spectrometry detector (GC/MS) by utilizing an internal standard method. The detecting method provided by the invention is capable of improving the repeatability of the measuring method, the purification method of the extract liquor is simple and effective, the pollution of a sample solution on a chromatographic column and an ion source can be reduced, the sensitivity is good, and the recovery is good.
Description
Technical field
The present invention relates to tobacco components assay method, be specifically related to the GC/MS assay method of humectant in a kind of cigarette shreds (being 1,2-PD, glycerine and triethylene glycol).
Background technology
In production of cigarettes, transport, storage, sale and sucking process, be to keep moisture content in leaves, increase pliability, reduce make broken, reduce pungency and improve jealously, need in tobacco, add humectant.1,2-PD, glycerine and triethylene glycol conventional humectant while being production of cigarettes, with all containing in feed liquid, significant to the Accurate Determining of these compositions in cigarette shreds at many flavouring essence for tobaccos and cigarette.The 9th article of the World Health Organization's Framework Convention on Tobacco Control (FCTC) and the 10th article of working group together with nicotine (nicotine) and ammoniac compounds, classify humectant as preferential tobacco and tobacco product three compounds of determining detection method.
The method of early detection tobacco humectant mainly contains titrimetry, hydrometer method, optical activity method, spectroscopic methodology etc.In recent years, gas chromatography (GC) method and high performance liquid chromatography (HPLC) method etc. start for detection of this type of polyol compound, tobacco business is also issued the assay method of humectant in tobacco and tobacco product: " tobacco and tobacco product 1,2-propylene glycol, the mensuration of glycerine: vapor-phase chromatography ", but these two kinds of method detection limits are conventionally higher.2010, Zhang Jie etc. have reported and have utilized GC/MS method to detect 1 in smoke-free tobacco product simultaneously, 2-propylene glycol, glycerine and triethylene glycol, but it is fairly simple that the weak point of this method is pre-treating method, sample extraction liquid, through further purifying, does not easily pollute chromatographic column and ion gun, thereby increase maintenance cost, reduces chromatographic column and ionogenic life-span.Moreover, due to the particularly high viscosity of glycerine of humectant, make its skewness in cigarette shreds, easily cause measurement result repeatability and the recovery bad.
Summary of the invention
Object of the present invention is improved for the deficiency of above-mentioned existing analytical technology just, specialized designs humectant 1,2-PD in a kind of cigarette shreds, the GC/MS assay method of glycerine and triethylene glycol.The detection method that this method provides, has improved the repeatability of measuring method, and sample liquid purification method is simply effective, can lower extract to chromatographic column and ionogenic pollution, and highly sensitive, and the recovery is good.
The object of the invention is to be achieved through the following technical solutions: humectant 1,2-PD in a kind of cigarette shreds, the GC/MS assay method of glycerine and triethylene glycol, dry, pulverize tobacco sample and sieves; Extract, respectively through disperseing after Solid-Phase Extraction (SPE) centrifuge tube and filter membrane purified treatment, by gas chromatography and mass detector (GC/MS), adopts inner mark method ration to detect the content of three kinds of humectants in pipe tobacco; Concrete steps are as follows:
1. mixing of cigarette shreds sample: tobacco sample is placed in to 40
oin C baking oven, after 1 hour, pipe tobacco is taken out, be placed in high speed disintegrator and pulverize 0.45 millimeter of aperture standard sieve of rear mistake 5-10 second;
2. the extraction of cigarette shreds sample: take the cigarette shreds powder of approximately 4 g balances after 48 hours, be placed in 100 mL conical flasks, adding 50 mL internal standard compound (1,3-BDO) concentration is the methanol extraction agent of 2 mg/mL, tool plug concussion extraction 1 h, after extract lucifuge is left standstill to 30 min.(internal standard compound also can be selected other material, as BDO)
3. the purification of extract: 2 mL extracts are placed in to dispersion Solid-Phase Extraction (SPE) centrifuge tube that contains 150 mg anhydrous magnesium sulfates and 25 mg N-propyl group ethylenediamines (PSA), on whirlpool mixed instrument with rotating speed vortex oscillation 2 min of 2000 rpm, with centrifugal 10 min of rotating speed of 10000 rpm, get after supernatant is crossed 0.22 μ m filter membrane and carry out GC/MS analysis again.
4. GC conditions:
Chromatographic column: 30 mm × 0.25, m × 0.25 μ m polyglycol elastic capillary pipe chromatographic columns (DB-WAX chromatographic column or other equivalent posts).
Injector temperature: 250
oc.
Carrier gas: helium (purity >=99.999%), constant current flow velocity: 1.0 mL/min.
Sample size: 1 μ L, split sampling, split ratio 100:1.
Heating schedule: initial temperature 90
oc, with 15
othe speed to 180 of C/min
oc, keeps 8 min, with 50
othe speed of C/min rises to 230
oc, keeps 5 minutes, and later operational mode is 250
ounder C condition, keep 10 min.
5. mass spectrum condition:
Transmission line temperature: 250
oc; Ionization mode: electron bombardment ionization source (EI); Ionizing energy: 70 eV; Ion source temperature 250
oc; Quadrupole rod temperature: 150
oc; Solvent delay: 3 min; Detecting pattern: select ion detection (SIM).
Compared with prior art, the present invention has following excellent results:
1. the inventive method is placed on tobacco sample baking and in high speed disintegrator, pulverizes and sieve in pretreatment process, makes the distribution of humectant between parallel sample more even.Experimental result shows, analysis result of the present invention is better than the sample repeatability without pulverizing and sieving and the recovery.
2. dispersion Solid-Phase Extraction (SPE) centrifuge tube through containing 150 mg anhydrous magnesium sulfates and 25 mg N-propyl group ethylenediamines (PSA) and the purification of 0.22 μ m filter membrane by extract in the inventive method sample pretreatment process, simple to operate, time saving and energy saving, reduce extract to chromatographic column and ionogenic pollution.
3. the inventive method has adopted the GC/MS of high sensitivity and strong anti-interference ability to detect, therefore have operation accurately, low, the recovery of detection limit and the advantage such as reproducible.
1) standard curve range of the inventive method, detection limit and quantitative limit:
This method, by joined standard operation solution least concentration solution replicate determination 10 times, is got respectively to 3 times of analysis result standard deviation and 10 times as its detection limit and quantitative limit, as
table 1shown in.
table 1. the detection limit of three kinds of humectants and quantitative limit in cigarette shreds
2) repeatability of the inventive method and recovery of standard addition:
3 parts, cigarette shreds sample getting known content, low according to 50%(respectively), in 100%() and 200%(high) three kinds of levels add 1,2-PD, glycerine and triethylene glycol standard items, 5 samples of the horizontal replication of each interpolation.Sample after mark-on carries out respectively pre-treatment and GC/MS to be analyzed, and by former content, add scalar and mark-on measured quantity calculate recovery rate, the results are shown in
table 2.From
table 2known, the average recovery of standard addition of 1,2-PD, glycerine, triethylene glycol is all between 97.4%-113.1%, and average relative standard deviation (RSD), between 1.7-6.2, shows that this method accuracy is high, reproducible, is applicable to quantitatively.
table 2. the recovery of standard addition of three kinds of humectants in cigarette sample pipe tobacco
Note: LOD is detectability
Brief description of the drawings
fig. 1for operational flowchart of the present invention
fig. 2for 1,2-PD, glycerine and triethylene glycol standard solution chromatogram
fig. 3for 1,2-PD, glycerine and triethylene glycol chromatogram in cigarette sample
Embodiment
The present invention is further described by following specific embodiment, but does not limit the present invention.
1. instrument, reagent and instrument condition of work
1) instrument
The mono-quadrupole rod gas chromatograph-mass spectrometer of DSQ II (Thermo Scientific company of the U.S.); AE163 electronic balance (sensibility reciprocal: 0.0001 g, Mettler company of Switzerland); HY-8 velocity-modulated oscillator (Changzhou Guohua Electric Appliance Co., Ltd.); High speed disintegrator (Wuhan Yin Cai Science and Technology Ltd.); The desk-top frozen type hydro-extractor of Germany's SIGMA 3-30K-high speed.
2) reagent
Methyl alcohol (chromatographically pure), 1,3-BDO (interior mark, CAS:107-88-0, purity >=99%), 1,2-propylene glycol (CAS:57-55-6, purity >=99.5%), glycerine (CAS:56-81-5, purity >=99.5%), triethylene glycol (CAS:112-27-6, purity >=99.5%), N-propyl group ethylenediamine adsorbent (PSA, analyze pure), anhydrous magnesium sulfate (analyzing pure).
3) instrument condition of work
A) GC conditions:
Chromatographic column: 30 mm × 0.25, m × 0.25 μ m polyglycol elastic capillary pipe chromatographic columns (DB-WAX chromatographic column or other equivalent posts)
Injector temperature: 250
oc.
Carrier gas: helium (purity >=99.999%), constant current flow velocity: 1.0 mL/min.
Sample size: 1 μ L, split sampling, split ratio 100:1.
Heating schedule: initial temperature 90
oc, with 15
othe speed to 180 of C/min
oc, keeps 8 min, with 50
othe speed of C/min rises to 230
oc, keeps 5 minutes, and later operational mode is 250
ounder C condition, keep 10 min.
B) mass spectrum condition:
Transmission line temperature: 250
oc; Ionization mode: electron bombardment ionization source (EI); Ionizing energy: 70 eV; Ion source temperature 250
oc; Quadrupole rod temperature: 150
oc; Solvent delay: 3 min; Detecting pattern: select ion detection (SIM).
C) qualitative and quantitative test
Adopt standard substance retention time, component characteristics ion to be measured, each qualitative abundance of ions ratio to carry out qualitative analysis.Adopt the fractional scanning of selectivity ion scan pattern, carry out quantitative test with characteristic ion.Each component characteristics ion is selected to see
table 3.
table 3. the quantitative and qualitative selection ion of humectant and internal standard compound table
| Sequence number | Compound title | Qualitative ion and abundance ratio thereof | Quota ion | Auxiliary quota ion |
| 1 | 1,2-PD | 45:43（100:20） | 45 | 43 |
| 2 | 1,3-BDO | 43:72（100:28） | 72 | 43 |
| 3 | Glycerine | 61:43（100:79） | 61 | 43 |
| 4 | Triethylene glycol | 45:89（100:19） | 45 | 89 |
2. determining of the preparation of standard operation solution and standard working curve
1) configuration of standard reserving solution
A) mark storing solution in: accurately take 20 g 1,3-BDOs in 100 mL volumetric flasks, to scale, being configured to concentration is the interior mark storing solution of 200mg/mL, 4 by methanol constant volume
ounder C condition, preserve.
B) extractant: accurately pipette mark storing solution in 20 mL and, in the volumetric flask of 2 L, in scale, be mixed with the extractant that 1,3-BDO concentration is 2 mg/mL by methanol constant volume.
C) 1,2-PD storing solution (50 mg/mL): take approximately 1.0 g(+/-0.05g) 1,2-PD is in 100 mL volumetric flasks, with extractant constant volume.4
ounder C condition, preserve.
D) glycerine storing solution (100 mg/mL): take approximately 10.0 g(+/-0.05 g) glycerine in 100 mL volumetric flasks, with extractant constant volume.4
ounder C condition, preserve.
E) triethylene glycol storing solution (25 mg/mL): take approximately 2.5 g(+/-0.05 g) triethylene glycol in 100 mL volumetric flasks, with extractant constant volume.4
ounder C condition, preserve.
2) preparation of mixed standard solution
Accurately pipette respectively 2.5 mL1,2-propylene glycol storing solution, 20 mL glycerine storing solutions and 16 mL triethylene glycol storing solutions are in 100 mL volumetric flasks, with extractant constant volume, prepare in mixed standard solution 1,2-propylene glycol concentration is 0.25 mg/mL, and glycerine concentration is 20 mg/mL, and triethylene glycol concentration is 4 mg/mL.4
ounder C condition, preserve.
3) preparation of standard operation solution
table 4. the preparation of standard operation solution
| Standard solution | Pipette mixed standard solution volume (mL) | Constant volume (mL) | 1,2-PD (mg/mL) | Glycerine (mg/mL) | Triethylene glycol (mg/mL) |
| 1 | 0.2 | 10 | 0.005 | 0.4 | 0.08 |
| 2 | 0.4 | 10 | 0.01 | 0.8 | 0.16 |
| 3 | 0.8 | 10 | 0.02 | 1.6 | 0.32 |
| 4 | 1.6 | 10 | 0.04 | 3.2 | 0.64 |
| 5 | 3.2 | 10 | 0.08 | 6.4 | 1.28 |
| 6 | 4.8 | 10 | 0.12 | 9.6 | 1.92 |
Note: institute's configuration standard working solution is with extractant constant volume.
4) determining of standard working curve
Working stamndard solution is carried out to chromatographic determination, calculate in each standard solution 1,2-propylene glycol, glycerine and triethylene glycol and interior target peak area ratio, make 1, the typical curve of 2-propylene glycol, glycerine and triethylene glycol concentration and peak area ratio or calculate regression equation, typical curve should be linear relationship, coefficient R
2should be not less than 0.995.Often carry out, after 20 sample determinations, should adding the working stamndard solution of an intermediate concentration, exceed 5% if the value measuring and initial value differ, should re-start the making of whole typical curve.
3. the pre-treatment of cigarette shreds sample and assay
1) tobacco sample is placed in to 40
oin C baking oven, after 1 hour, pipe tobacco is taken out, be placed in high speed disintegrator and pulverize 0.45 millimeter of aperture standard sieve of rear mistake 5-10 second;
2) take the cigarette shreds powder of approximately 4 g balances after 48 hours, be placed in 100 mL conical flasks, adding internal standard compound (1,3-BDO) concentration is the methanol extraction agent of 2 mg/mL, tool plug concussion extraction 1 h, after extract lucifuge is left standstill to 30 min.
3) 2 mL extracts are placed in to 2 mL that contain 150 mg anhydrous magnesium sulfates and 25 mg N-propyl group ethylenediamines (PSA) and disperse Solid-Phase Extraction (SPE) centrifuge tube, on whirlpool mixed instrument with rotating speed vortex oscillation 2 min of 2000 rpm, with centrifugal 10 min of rotating speed of 10000 rpm, get after supernatant is crossed 0.22 μ m filter membrane and carry out GC/MS analysis again.
4) according to instrument test condition working sample, twice of each sample replication.
5) in sample the content of 1,2-PD, glycerine and triethylene glycol with shown in following formula:
In formula: humectant content in M-sample pipe tobacco, unit is mg/g
The concentration of 1,2-PD, glycerine and triethylene glycol in C-extraction solution, unit is every milliliter (mg/mL) of milligram
The volume of V-extract, unit is milliliter (mL)
The quality of m-sample, unit is gram (g)
Taking the mean value of twice replicate determination as final measurement result.
table 5.the testing result of three kinds of cigarette shreds samples
| Cigarette shreds sample | 1,2-PD (mg/g) | Glycerine (mg/g) | Triethylene glycol (mg/g) |
| 1 | 0.550 | 20.100 | < LOD |
| 2 | 0.250 | 22.700 | < LOD |
| 3 | 0.458 | 2.703 | < LOD |
Note: LOD is detectability
Claims (1)
1. a humectant 1,2-PD in cigarette shreds, the GC/MS assay method of glycerine and triethylene glycol, is characterized in that: tobacco sample be dry, pulverize and sieve; Extract, respectively through disperseing after Solid-Phase Extraction (SPE) centrifuge tube and filter membrane purified treatment, by gas chromatography and mass detector (GC/MS), adopts inner mark method ration to detect the content of three kinds of humectants in pipe tobacco; Concrete steps are as follows:
1) mixing of cigarette shreds sample: tobacco sample is placed in to 40
oin C baking oven, after 1 hour, pipe tobacco is taken out, after pulverizing, cross 0.45 millimeter of aperture standard sieve;
2) extraction of cigarette shreds sample: take the cigarette shreds powder of approximately 4 g balances after 48 hours, be placed in 100 mL conical flasks, add the methanol extraction agent that contains internal standard compound, tool plug concussion extraction 1 h, after extract lucifuge is left standstill to 30 min; Described internal standard compound is 1,3-BDO or BDO, and the concentration of internal standard compound in methanol extraction liquid is 2 mg/mL;
3) purification of extract: 2 mL extracts are placed in to dispersion Solid-Phase Extraction (SPE) centrifuge tube that contains 150 mg anhydrous magnesium sulfates and 25 mg N-propyl group ethylenediamines (PSA), on whirlpool mixed instrument with rotating speed vortex oscillation 2 min of 2000 rpm, with centrifugal 10 min of rotating speed of 10000 rpm, get after supernatant is crossed 0.22 μ m filter membrane and carry out GC/MS analysis again;
4) GC conditions:
Chromatographic column: 30 mm × 0.25, m × 0.25 μ m polyglycol elastic capillary pipe chromatographic columns;
Injector temperature: 250
oc,
Carrier gas: helium, constant current flow velocity: 1.0 mL/min;
Sample size: 1 μ L, split sampling, split ratio 100:1;
Heating schedule: initial temperature 90
oc, with 15
othe speed to 180 of C/min
oc, keeps 8 min, then with 50
othe speed of C/min rises to 230
oc, keeps 5 minutes, and later operational mode is 250
ounder C condition, keep 10 min;
5) mass spectrum condition:
Transmission line temperature: 250
oc; Ionization mode: electron bombardment ionization source (EI); Ionizing energy: 70 eV; Ion source temperature: 250
oc; Quadrupole rod temperature: 150
oc; Solvent delay: 3 min; Detecting pattern: select ion detection (SIM).
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3618213A (en) * | 1970-04-27 | 1971-11-09 | Nat Patent Dev Corp | Denture liners |
| CN101963604A (en) * | 2010-10-29 | 2011-02-02 | 川渝中烟工业公司 | Method for measuring sterol in tobaccos |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6818231B2 (en) * | 2001-12-12 | 2004-11-16 | Brian Alexis | Treatment of vulvovaginitis with spirostanol enriched extract from Tribulus terrestris |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3618213A (en) * | 1970-04-27 | 1971-11-09 | Nat Patent Dev Corp | Denture liners |
| CN101963604A (en) * | 2010-10-29 | 2011-02-02 | 川渝中烟工业公司 | Method for measuring sterol in tobaccos |
Non-Patent Citations (4)
| Title |
|---|
| GC/MS法同时检测无烟气烟草制品中的1,2-丙二醇、丙三醇和三甘醇;张杰等;《烟草科技》;20110320(第3期);第37～38页 * |
| 严会会等.高效液相色谱串联质谱法分析烟草中15中农药残留.《烟草科技》.2011,(第7期), |
| 张杰等.GC/MS法同时检测无烟气烟草制品中的1 2-丙二醇、丙三醇和三甘醇.《烟草科技》.2011 |
| 高效液相色谱串联质谱法分析烟草中15中农药残留;严会会等;《烟草科技》;20110720(第7期);第44页 * |
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