CN105738528A - GC-MS/MS analysis method for dinitraniline pesticide residues in mainstream cigarette smoke particulate matter - Google Patents

Abstract

The invention discloses a GC-MS/MS analysis method for dinitraniline pesticide residues in mainstream cigarette smoke particulate matter.The method comprises the following steps that 1, a pesticide standard substance solution is prepared, determination is performed through a gas chromatography-tandem mass spectrum, and fitting is performed to obtain a standard curve; 2, a Cambridge filter trapping the mainstream cigarette smoke particulate matter is vibrated and extracted through acetonitrile, an extracting solution is subjected to dispersive liquid-liquid microextraction to perform centrifugal treatment, and water and carbon tetrachloride are added into the extracting solution to be subjected to dispersive liquid-liquid microextraction, wherein the addition volume size of water to carbon tetrachloride is 1:3-5:0.1-0.3; 3, the centrifuged precipitation facies is determined on the condition of the gas chromatography-tandem mass spectrum, and the determined result is substituted into the standard curve to obtain the content of pesticide residues in the mainstream cigarette smoke particulate matter.The dispersive liquid-liquid microextraction technology and the GC-MS/MS are combined, and the sample processing step is simplified, so that the detection result is more accurate and stable, and the pesticide residue situation in reaction flue gas can be stably and objectively reflected.

Description

In cigarette mainstream smoke condensate, the GC-MS/MS of dinitroaniline pesticide residue analyzes method

Technical field

The invention belongs to cigarette smoke the Detection Technologies of Pesticide Residues field, relate generally to cigarette mainstream flue gas The determination techniques of dinitroaniline pesticide residue in granule phase substance, a kind of cigarette mainstream flue gas In granule phase substance, the GC-MS/MS of dinitroaniline pesticide residue analyzes method.

Background technology

Nicotiana tabacum L. is during plantation, production and processing and storage, it is to avoid the not use of pesticide, thus makes Become relevant pesticide residual on Nicotiana tabacum L., Medicated cigarette have again during burning and sucking part pesticide through aspirate into Enter in main flume.Wan Yilun etc. (Wan Yilun, Zhang Hongfei, Gao Chuanchuan, etc. 126 kinds of agricultures in Medicated cigarette Medicine transfer behavior [J] during burning and sucking. Guizhou Agricultural Sciences, 2015 (05): 101-106.) research table In bright tobacco shred, 126 kinds of pesticide average mobility in cigarette mainstream flue gas is 7.1%, the most different Di Family name's agent reaches as high as 31.7%, and the transfer behavior that variety classes pesticide is during Medicated cigarette burns and sucks is poor Different the biggest, dinitroaniline pesticide mobility in main flume is 8.67%～13.16%.

The detection method of dinitroaniline pesticide residue analysis in cigarette smoke mainly there is gas phase color Spectrometry, gas chromatography mass spectrometry method etc., pretreatment process mainly has mechanical shaking extraction, accelerated solvent extraction etc., mirror Persticide residue in total particulate matter in mainstream smoke is relatively low, therefore in pre-treatment operating process it is generally required to Extract is carried out concentration and is beneficial to detection analysis, but this allows pre-treatment become loaded down with trivial details, Er Qienong simultaneously Compression process may result in the response rate of part pesticide to be reduced.

The Chinese patent literature of Publication No. CN101393183A discloses in Nicotiana tabacum L. and tobacco product The assay method of six kinds of dinitroaniline persticide residues, extraction in accelerated solvent extraction (ASE) Take pesticide residues, directly measure Nicotiana tabacum L. and Nicotiana tabacum L. with gas chromatogram (joining electron capture detector (ECD) (ECD)) Goods stop bud element, flumetralim, isopropalin, pendimethalin, benfluralin and trefanocide etc. six kinds The method of dinitroaniline persticide residue.

The Chinese patent literature of Publication No. CN103293266A discloses a kind of analysis Nicotiana tabacum L. and cigarette The GC-MS/MS method of dinitroaniline pesticide residue in straw-made articles, by sample after water infiltration, Add acetonitrile and internal standard substance, then extraction of ocean eddies, layering of saltouing, afterwards with sodium-chloride water solution and just Hexane carries out secondary and saltouts and liquid-liquid extraction, takes normal hexane supernatant, with gas chromatography tandem mass spectrometry The method that method is measured.The method pre-treatment is comparatively laborious.

Persticide residue in total particulate matter in mainstream smoke is relatively low, it is generally required to right in pre-treatment operating process Extract carries out concentration and is beneficial to detection analysis, but loaded down with trivial details pre-treatment may result in part pesticide The response rate reduces, and testing result is inaccurate.

Summary of the invention

The purpose of the present invention aims to provide a kind of filter disc warp that trapping has cigarette mainstream smoke condensate The side that after extraction, dispersive liquid-liquid microextraction, gas chromatography tandem mass spectrometry (GC-MS/MS) is measured Method.The method can quickly, accurately detect the Multiple Pesticides residual in cigarette mainstream smoke condensate, surveys Determine result accurate, reproducible.

In a kind of cigarette mainstream smoke condensate, the GC-MS/MS of dinitroaniline pesticide residue divides Analysis method, comprises the following steps:

(1): preparation standard sample of pesticide solution, by gas chromatography tandem mass spectrometry (GC-MS/MS) It is measured and matching obtains standard curve；

(2): trapping is had the cambridge filter of cigarette mainstream smoke condensate through acetonitrile mechanical shaking extraction, Extracting solution carries out centrifugal treating after being dispersed through liquid-liquid micro-extraction；

In extracting solution, add water and carbon tetrachloride carry out dispersive liquid-liquid microextraction, wherein, extracting solution: Water: the volume ratio that adds of carbon tetrachloride is 1:3-5:0.1-0.3；

(3): the sedimentary facies after Li Xin is again under the conditions of the gas chromatography tandem mass spectrometry of step (1) It is measured；And measurement result substitutes into standard curve, and to try to achieve cigarette mainstream smoke condensate Pesticides residual Stay content.

The pesticide variety that may remain in cigarette mainstream flue gas is a lot, and content is relatively low, it is achieved in flue gas The detection difficulty of the high sensitivity of pesticide is bigger.The present invention uses dispersive liquid-liquid microextraction technology to Medicated cigarette Main flume pesticide residues composition extracts, and instead of traditional nitrogen and the concentration step such as blows, simplifies Sample handling procedure.Additionally, dispersive liquid-liquid microextraction technology is combined with GC-MS/MS, can Testing result is made to react flue gas Pesticide Residues situation more accurately, stably, objectively, it is achieved that Medicated cigarette Detect while 6 kinds of dinitroaniline pesticide residues of total particulate matter in mainstream smoke, improve detection efficiency.

In step (1) and step (3), using internal standard method to be measured, the internal standard substance of interpolation is three Phenyl phosphate ester.

As, during preparing standard solution and testing sample solution (sedimentary facies), add inner mark solution. Described inner mark solution is the acetonitrile solution of triphenyl.As, concentration is the TPP of 20 μ g/mL The acetonitrile solution of (triphenyl).

In step (1), preparation standard sample solution processes is:

Weigh standard sample of pesticide respectively, with acetone solution, then add internal standard substance by dilution in acetonitrile extremely There is the standard solution containing various pesticide of Concentraton gradient；

Described standard sample of pesticide solution comprises benfluralin, butralin, flumetralim, isopropalin, Pendimethalin and trefanocide.

As, the manner of formulation of standard solution is: weigh 0.01g (being accurate to 0.1mg) respectively described Various standard sample of pesticide, be placed in same 10mL volumetric flask, obtain 1 with acetone solution constant volume The mixed standard solution I of mg/mL；Accurately pipette 0.2mL mixed standard solution I in 100mL In volumetric flask, use acetonitrile constant volume, be configured to the mixed standard solution II that each pesticide concentration is 2 μ g/mL； Accurately pipette 0.01mL, 0.02mL, 0.05mL, 0.1mL, 0.2mL, 0.5mL and 1mL Mixed standard solution II, be placed in 10mL volumetric flask, accurately add 100 μ L inner mark solutions (dense Degree is the acetonitrile solution of the TPP of 20 μ g/mL), use acetonitrile constant volume, obtain series standard working solution. Preparation series standard solution concentration be: 2ng/mL, 4ng/mL, 10ng/mL, 20ng/mL, 40ng/mL、100ng/mL、200ng/mL。

After standard solution has been prepared, when carrying out GC-MS/MS mensuration, GC realizes composition in sample Separation.MS realizes the qualitative and quantitative analysis of composition in sample.The detection parameter pair of GC and MS The impact of the separating degree of testing sample, detection limit is the biggest.

In the present invention, in step (1) and step (3), when GC-MS/MS measures, use Chromatographic condition be:

Chromatographic column: HP-5MS quartz capillary column: 30m × 0.25mm × 0.25 μm；

Injector temperature: 280 DEG C；Temperature programming: initial temperature 60 DEG C, keep after 1min with 40 DEG C/min rises to 170 DEG C, then rises to 310 DEG C with 10 DEG C/min, keeps 3min；Program after post: 310 DEG C, keep 5min；

Splitless injecting samples, does not shunt time 1min；

Carrier gas: helium, purity is 99.999%；

Constant current mode, flow velocity is 1.2mL/min；Sample size: 1 μ L.

The Mass Spectrometry Conditions used is:

Ionization energy 70eV；Heater current: 35 μ A；Ion source temperature: 300 DEG C；Quadrupole rod temperature: 180℃；Transmission line temperature: 280 DEG C；Q2 collides gas: nitrogen, purity 99.999%, flow 1.5 mL/min；Quenching gas: helium, purity 99.999%, flow 2.25mL/min；Solvent delay 5 min；Scan mode: multiple-reaction monitoring.

In step (1), standard solution is after GC-MS/MS analyzes, then with pesticide and internal standard substance Two grades select its respective concentration of ion peak areas comparison to carry out regression analysis, obtain standard curve.

The drawing process (step (1)) of step (2) (i.e. fume sample pre-treatment) and standard curve is Parallel step, without sequencing.

In step (2), the extracting method of main flume Pesticide Residues will directly influence test knot The accuracy of fruit and credibility.In the present invention, collected the granule phase substance of main flume by cambridge filter, Extract as dispersant with acetonitrile again.In extracting solution, add water and extractant again, fully shake, Be centrifuged subsequently, will the granule phase substance enrichment of main flume in extractant, then to the extractant after centrifugal Layer is measured.

Every cambridge filter is collected 3-5 by ISO 3308:2000 method and is propped up the main flume of cigarette sample Granule phase substance.During mechanical shaking extraction, every cambridge filter adds the acetonitrile of 20-30mL.

As preferably, mechanical shaking extraction process is carried out on shaker, and the frequency of oscillation of shaker is 150-200r/min, the oscillation extraction time is 20-40min.Further preferably, the oscillation frequency of shaker Rate is 150r/min, and the oscillation extraction time is 30min.

As preferably, in dispersive liquid-liquid microextraction, extracting solution: water: carbon tetrachloride add volume ratio For 1:5:0.2.

The extractant that the present invention uses is carbon tetrachloride, the organic principle that can be fully enriched with in extracting solution, Make testing result more objective.

As preferably, in centrifugal process, centrifugal rotational speed is 5000-8000r/min, and centrifugation time is 3-5min.Further preferably, in centrifugal process, centrifugal rotational speed is 5000r/min, and centrifugation time is 3min。

As, the preparation method (sample pretreatment process) of cigarette mainstream smoke condensate testing sample is: Collect the total particulate matter in mainstream smoke of 4 cigarette samples by ISO 3308:2000 method after, take off sword Bridge filter disc；(inner mark solution, concentration is the TPP of 20 μ g/mL accurately to add 0.1mL internal standard working solution Acetonitrile solution) and 20mL acetonitrile in conical flask, with 150r/min oscillation extraction 30min；Take 1mL acetonitrile extraction liquid, in 10mL tool plug conical centrifuge tube, adds 5mL water, adds 200 μ L carbon tetrachloride, continues vibration 30s, forms uniform emulsion, be centrifuged 3min with 5000r/min After, by syringe suction foot organic solution in chromatograph bottle, carry out step (3) subsequently GC-MS/MS analyzes.

The chromatographic condition of step (3) and Mass Spectrometry Conditions are equal to step (1).

Above-mentioned chromatographic condition and Mass Spectrometry Conditions highly sensitive, capacity of resisting disturbance is strong, simple to operate, Reproducible, can realize cigarette mainstream smoke condensate is examined while dinitroaniline pesticide residue Survey, improve detection efficiency.The detection limit of the present invention is low, in terms of 10 times of signal to noise ratios (S/N=10) Calculating quantitative limit (LOQ), quantitative detection limit is between 0.2ng/cig～1.0ng/cig.Side of the present invention The response rate of method is high, reproducible, wherein, the response rate of pesticide between 71.4%～118.5%, Relative standard deviation (RSD) is between 1.9%～8.3%.

The method of the present invention instead of traditional nitrogen blow etc. dense by introducing dispersive liquid-liquid microextraction technology Contracting step, simplifies existing sample treatment, and is provided that the most more stable measurement result. Compared with prior art the inventive method has a following excellent results:

(1) the inventive method has been firstly introduced dispersive liquid-liquid microextraction technology and has been applied to cigarette mainstream flue gas Dinitroaniline pesticide residues quantitative analysis in granule phase substance, simplifies sample pre-treatments, concentration effect Significantly.

(2) the inventive method have employed the GC-MS/MS of high sensitivity and strong anti-interference ability and examines Survey, therefore there is the advantage that operation is accurate, highly sensitive and reproducible.

(3) the inventive method achieves 6 kinds of dinitroaniline pesticide residues of cigarette mainstream smoke condensate While detect, improve detection efficiency.

Detailed description of the invention

Following example are pressed aforesaid operations method and are implemented:

Reagent:

Standard sample of pesticide: benfluralin, butralin, flumetralim, isopropalin, pendimethalin, fluorine Happy spirit；

Internal standard substance: TPP, for standard substance.The acetonitrile being configured to the TPP that concentration is 20 μ g/mL is molten Liquid (inner mark solution, internal standard working solution)；

Acetonitrile, acetone, carbon tetrachloride, be chromatographically pure reagent.

Pure water, meets the requirement of one-level water in GB/T 6682.

Instrument:

U.S.'s Agilent 7000B gas chromatogram-triple quadrupole bar tandem mass spectrometer (GC-MS/MS)； Britain's Si Rulin SM 450 linear type smoking machine；U.S.'s Labnet VtexMixer VX200 agitator； Germany's Sigma High speed refrigerated centrifuge.

Embodiment 1:

Step (1): preparation pesticide standard solution:

Weigh 0.01g (being accurate to 0.1mg) various standard sample of pesticide, be placed in same 10mL capacity In Ping, obtain the mixed standard solution I of 1mg/mL with acetone solution constant volume；Accurately pipette 0.2 ML mixed standard solution I, in 100mL volumetric flask, uses acetonitrile constant volume, is configured to each pesticide concentration It it is the mixed standard solution II of 2 μ g/mL；Accurately pipette 0.01mL, 0.02mL, 0.05mL, The mixed standard solution II of 0.1mL, 0.2mL, 0.5mL and 1mL, is placed in 10mL volumetric flask In, accurately add 100 μ L inner mark solutions, use acetonitrile constant volume, obtain series standard working solution. Preparation series standard solution concentration be: 2ng/mL, 4ng/mL, 10ng/mL, 20ng/mL, 40ng/mL、100ng/mL、200ng/mL。

The standard working solution of the variable concentrations prepared is injected GC-MS/MS, carries out with internal standard method The quantitative analysis of pesticide, when GC-MS/MS measures, the chromatographic condition of employing is: chromatographic column: HP-5MS quartz capillary column (30m × 0.25mm × 0.25 μm)；Injector temperature: 280℃；Temperature programming: initial temperature 60 DEG C, rises to 170 DEG C with 40 DEG C/min after keeping 1min, Rise to 310 DEG C with 10 DEG C/min again, keep 3min；Program after post: 310 DEG C, keeps 5min； Splitless injecting samples, does not shunt time 1min；Carrier gas: helium (purity is 99.999%)；Constant current Pattern, flow velocity is 1.2mL/min；Sample size: 1 μ L.Total run time is 20.75min.

The Mass Spectrometry Conditions used is: ionization energy 70eV；Heater current: 35 μ A；Ion source temperature: 300℃；Quadrupole rod temperature: 180 DEG C；Transmission line temperature: 280 DEG C；Q2 collides gas: nitrogen is (pure Degree 99.999%, flow 1.5mL/min), quenching gas helium (purity 99.999%, flow 2.25 mL/min)；Solvent delay 5min；Scan mode: multiple-reaction monitoring (MRM).Multiple agriculture Medicine and interior target retention time and mass spectrometry parameters are shown in Table 1.

Table 1

Its respective concentration of ion peak areas comparison is selected to carry out regression analysis with pesticide and interior target two grades, Obtain standard curve, be shown in Table 2.

Detection limit:

In blank extracting solution, add the standard solution of embodiment 1, and inject GC-MS/MS analysis, Calculating quantitative limit (LOQ) with 10 times of signal to noise ratios (S/N=10), quantitative detection limit is at 0.2ng/cig ～between 1.0ng/cig, data are shown in Table 2.

Repeatability and recovery of standard addition:

Standard sample of pesticide solution (concentration is 100ng/mL) is added, then in blank filter disc sample Carry out pre-treatment (step (2)) respectively and GC-MS/MS analyzes, and measure according to adding scalar sum Value calculates its response rate, the results are shown in Table 2 and (linear equation of pesticide, regression coefficient, quantitative limit, returns Yield and repeatability (n=6)).

Table 2

As can be seen from Table 2, the response rate of pesticide is between 71.4%～118.5%, and relative standard is inclined Difference (RSD), between 1.9%～8.3%, illustrates that the response rate of the inventive method is high, reproducible.

Step (2): sample treatment (sample pre-treatments):

Collect the total particulate matter in mainstream smoke of 4 cigarette sample A by ISO 3308:2000 method after, Take off cambridge filter；Accurately add 0.1mL internal standard working solution (acetonitrile solution of the TPP of 20 μ g/mL) With 20mL acetonitrile in conical flask, with 150r/min oscillation extraction 30min；Take 1mL acetonitrile to carry Take liquid and fill in conical centrifuge tube in 10mL tool, add 5mL water, add 200 μ L carbon tetrachloride, Continue vibration 30s, form uniform emulsion, after being centrifuged 3min with 5000r/min, with injection Device suction foot organic solution in chromatograph bottle, analysis to be detected.

Step (3): GC-MS/MS measures:

Sample after extracting is measured, records detection pesticide and select quasi-molecular ions face with interior target two grades Long-pending ratio, substitutes into standard curve, tries to achieve the content of various pesticide in sample A total particulate matter in mainstream smoke and be shown in Table 3。

Table 3

Note: " " expression does not detects

Embodiment 2:

As described in Example 1, select another cigarette sample B, record its total particulate matter in mainstream smoke middle peasant The content of medicine object is shown in Table 4.

Table 4

Note: " " expression does not detects

Embodiment 3:

As described in Example 1, select another cigarette sample C, record its total particulate matter in mainstream smoke middle peasant The content of medicine object is shown in Table 5.

Table 5

Note: " " expression does not detects

Being found by each embodiment, this method can quickly, accurately detect in cigarette mainstream smoke condensate Multiple Pesticides remains, and measurement result is accurate, reproducible；Dinitroaniline pesticide residue can be realized While detect, detection efficiency is high.

Comparative example 1:

It is with embodiment 1 difference, in step (2), acetonitrile extraction liquid: water: carbon tetrachloride Adding volume ratio is 2:5:0.2.In cigarette sample A total particulate matter in mainstream smoke, the content of various pesticide is shown in Table 6。

Table 6

Note: " " expression does not detects

Comparing with embodiment 1, as shown in Table 6, during improve split-phase liquid-liquid micro-extraction, acetonitrile carries Take the ratio of liquid, butralin in the cigarette sample A total particulate matter in mainstream smoke of embodiment 1, flumetralim, Pendimethalin does not all detect, and pesticide residues missing inspection situation occurs.This is primarily due at dispersant acetonitrile When ratio is excessive, pesticide object dissolubility in aqueous phase is greatly increased, thus has had a strong impact on point Dissipate the extraction efficiency of liquid-liquid micro-extraction process.

Comparative example 2:

It is with embodiment 1 difference, in step (2), uses dichloromethane to replace carbon tetrachloride to make For extractant, wherein, acetonitrile extraction liquid: water: the volume ratio that adds of dichloromethane keeps constant (1:5:0.2)。

Result shows, has no the layering of aqueous phase and organic facies during dispersive liquid-liquid microextraction after being centrifuged, Illustrate that dichloromethane can not reach good extract and separate effect under this ratio.

Claims (8)

1. the GC-MS/MS of dinitroaniline pesticide residue in a cigarette mainstream smoke condensate Analysis method, it is characterised in that comprise the following steps:

(1): preparation standard sample of pesticide solution, by gas chromatography tandem mass spectrometry (GC-MS/MS) It is measured and matching obtains standard curve；

(2): trapping is had the cambridge filter of cigarette mainstream smoke condensate through acetonitrile mechanical shaking extraction, Extracting solution carries out centrifugal treating after being dispersed through liquid-liquid micro-extraction；

In extracting solution, add water and carbon tetrachloride carry out dispersive liquid-liquid microextraction, wherein, extracting solution: Water: the volume ratio that adds of carbon tetrachloride is 1:3-5:0.1-0.3；

(3): the sedimentary facies after Li Xin is again under the conditions of the gas chromatography tandem mass spectrometry of step (1) It is measured；And measurement result substitutes into standard curve, and to try to achieve cigarette mainstream smoke condensate Pesticides residual Stay content.

2. in cigarette mainstream smoke condensate as claimed in claim 1, dinitroaniline pesticide is residual The GC-MS/MS stayed analyzes method, it is characterised in that in step (1) and step (3), adopt Being measured with internal standard method, the internal standard substance of interpolation is triphenyl.

3. in cigarette mainstream smoke condensate as claimed in claim 2, dinitroaniline pesticide is residual The GC-MS/MS stayed analyzes method, it is characterised in that in step (1), standard solution warp After GC-MS/MS analyzes, then it is corresponding with two grades of selection ion peak areas comparisons of pesticide and internal standard substance Concentration carries out regression analysis, obtains standard curve.

4. in cigarette mainstream smoke condensate as claimed in claim 1, dinitroaniline pesticide is residual The GC-MS/MS stayed analyzes method, it is characterised in that mechanical shaking extraction process is carried out on shaker, The frequency of oscillation of shaker is 150-200r/min, and the oscillation extraction time is 20-40min.

5. in cigarette mainstream smoke condensate as claimed in claim 1, dinitroaniline pesticide is residual The GC-MS/MS stayed analyzes method, it is characterised in that in centrifugal process, centrifugal rotational speed is 5000-8000r/min, centrifugation time is 3-5min.

6. in cigarette mainstream smoke condensate as claimed in claim 1, dinitroaniline pesticide is residual The GC-MS/MS stayed analyzes method, it is characterised in that in step (1) and step (3), When GC-MS/MS measures, the chromatographic condition of employing is:

Chromatographic column: HP-5MS quartz capillary column: 30m × 0.25mm × 0.25 μm；

Injector temperature: 280 DEG C；Temperature programming: initial temperature 60 DEG C, keep after 1min with 40 DEG C/min rises to 170 DEG C, then rises to 310 DEG C with 10 DEG C/min, keeps 3min；Program after post: 310 DEG C, keep 5min；

Splitless injecting samples, does not shunt time 1min；

Carrier gas: helium, purity is 99.999%；

Constant current mode, flow velocity is 1.2mL/min；Sample size: 1 μ L.

7. in cigarette mainstream smoke condensate as claimed in claim 6, dinitroaniline pesticide is residual The GC-MS/MS stayed analyzes method, it is characterised in that the Mass Spectrometry Conditions of employing is:

Ionization energy 70eV；Heater current: 35 μ A；Ion source temperature: 300 DEG C；Quadrupole rod temperature: 180℃；Transmission line temperature: 280 DEG C；Q2 collides gas: nitrogen, purity 99.999%, flow 1.5 mL/min；Quenching gas: helium, purity 99.999%, flow 2.25mL/min；Solvent delay 5 min；Scan mode: multiple-reaction monitoring.

8. in cigarette mainstream smoke condensate as claimed in claim 1, dinitroaniline pesticide is residual The GC-MS/MS stayed analyzes method, it is characterised in that comprise in described standard sample of pesticide solution Benfluralin, butralin, flumetralim, isopropalin, pendimethalin and trefanocide.