CN102590421A - Method for detecting musk xylene in tobacco additive - Google Patents

Abstract

The invention relates to a method for detecting musk xylene in a tobacco additive and belongs to the technical field of detection of tobacco additives. The method comprises the following steps of: weighing 1.0g of tobacco additive and putting into a 50mL conical flask with a plug, adding 10mL of ultrapure water of which electrical resistivity is 18.2 M Omega and dispersing a sample, adding 0.3mL of D15-musk xylene interior label solution with the concentration of 1mu g/mL, and 10mL of cyclohexane, oscillating and extracting at the temperature of 25 DEG C for 10 minutes, centrifuging a mixed solution at rotation speed of 4,000r/min by a centrifugal machine for 10 minutes, extracting an upper-layer organic phase, adding 10mL of chromatographically pure cyclohexane into a water phase of which the upper-layer organic phase is extracted, performing secondary extraction under the same operation conditions, centrifuging, and extracting an upper-layer organic phase; and mixing the organic phases extracted twice, drying by using anhydrous sodium sulfate, and metering the volume to ensure that the volume is 1mL for gas chromatography-mass spectrometer (GC-MC) determination. The method has the advantages of high sensitivity and capacity of effectively determining the content of musk xylene in the tobacco additive, and can determine the content of the musk xylene accurately and quickly, and is suitable for monitoring and self-inspection work of the highly-concerned musk xylene in the tobacco additive.

Description

The detection method of musk xylene in a kind of cigarette additive

Technical field:

The present invention relates to the detection method of musk xylene in a kind of cigarette additive, belong to cigarette additive detection technique field.

Background technology:

Musk xylene (Musk xylene) is a kind of nitroaromatic Moschus compound of synthetic, chemical name 2,4,6-trinitro--1,3-dimethyl-5-tert-butyl benzene.Characteristics such as it is special to have fragrance, and lasting is lasting, the fixastive of making cosmetics and fragrance for detergents commonly used ^[1]But because musk xylene has the report that can make mouse carcinogenic ^[2], and, classified as the high material of paying close attention to by chemicals management administration of European Union as high persistence, high biological accumulation property material ^[3,4]In July, 2009, international daily use chemicals fragrance association announced that also the P-xylene Moschus forbids comprehensively ^[5], domestic tobacco business also clearly is defined in and bans use of musk xylene in the cigarette additive.

At present, the research report of relevant musk xylene detection method mainly concentrates on cosmetics both at home and abroad ^[6-10], biological sample ^[9], environmental sample ^[12-15]Aspect, the method that relates to mainly comprise thin-layered chromatography (TLC) ^[6], gas chromatography-electron capture detection method (GC-ECD) ^[12], GC-MS (GC-MS) ^[7-9,14-15], gas chromatography-tandem mass spectrometry (GC-MS/MS) ^[10-11], nuclear magnetic resonance method (NMR) ^[13]

Through the document retrieval, do not see the public reported of the detection method of musk xylene in the cigarette additive.

List of references:

[1]Chen?S?N.Encyclopedia?of?Chinese?Chem?ical?Products.Beijing：Chemical?Industry?Press，1998.

[2] Harris S W. Shu Hongfu translates. the latest survey of the nitro musk in the daily chemical essence product: FDA [J]. and fragrance flavor and cosmetic, 1997, (3): 39-41.

[3] Liu Shuwen. synthetic perfume technical manual (second edition) [M]. Beijing: China Light Industry Press, 2009.

[4] captain Yu, Li Huailin. European Union's cosmetics-laws and regulations on the management and detection method and guide (second edition) [M]. Beijing: China Light Industry Press, 2010.

[5]IFRA?Newsletter?No.13，April?2009.http://www.ifraorg.Org

[6]Goh?C?L，Kwok?S?F.A?simple?method?of?qualitative?analysis?for?musk?ambrette，musk?ketone?and?musk?xylene?in?cologne[J].Contact?Dermatitis，1986，14：53-56.

[7]Mitjans?D，Ventura?F.Determination?of?fragrancesat?ng/L?levels?using?CLSA?and?GC/MS?detection[J].Water?Sci.Technol.，2005，52(10-11)：145-150.

[8] Luo Chuan, Wu Beilei, Yuan Lifeng. the high mensuration-GC-MS [J] of paying close attention to material-musk xylene of REACH in the essential wind oil. inspection and quarantine academic periodical, 2011,4:32-35/73.

[9] Yang Run .GC-MS method is measured the method [J] that three-type-person in the cosmetics makes Moschus. Chinese sanitary inspection magazine, 2003,13 (4): 456-457.

[10] horse is strong, white birch, Wang Chao, Ma Wei, Zhang Qing, Xiao Haiqing, Zhou Xin, Dong Yiyang, Wang Baolin. and SPE-isotopic dilution-gas chromatography-tandem mass spectrometry is measured the musk xylene [J] in the cosmetics. analytical chemistry, 2009,37 (12): 1776-1780.

[11]Mottaleb?M?A，Usenko?S，O’Donnelld?J?G，Ramirez?A?J，Brooks?B?W，Chambliss?C?K.Gas?chromatography-mass?spectrometry?screening?methods?for?select?UV?filters，synthetic?musks，alkylphenols，an?antimicrobial?agent，and?an?insect?repellentin?fish[J].J.Chromatogr.A，2009，1216：815-823.

[12]Polo?M，Garcia-Jares?C，LIompart?M.Optimization?of?a?sensitive?method?for?the?determination?of?nitro?musk?fragrances?in?waters?by?solid-phase?microextraction?and?gas?chromatography?with?micro?electron?capture?detection?using?factorial?experimental?design[J].Anal.Bioanal.Chem.，2007，388：1789-1798.

[13]Berset?J?D.Analysis?of?nitro-musk?compounds?and?their?amino?metabolites?in?liquid?sewage?sludges?using?NMR?and?mass?spectrometry[J].Anal.Chem.，2000，72：2124-2131

[14]Ramírez?N，MarcéR?M，Borrull?F.Development?of?a?stir?bar?sorptive?extraction?and?thermal?desorption-gas?chromatography-mass?spectrometry?method?for?determining?synthetic?musks?in?water?samples[J].J.Chromatogr.A，2011，1218：156-161.

[15]Kallenborn?R，Gatermann?R，Planting?S，Rimkus?G?G，Lund?M，Schlabach?M，Burkow?I?C.Gas?chromatographic?determination?of?synthetic?musk?compounds?in?Norwegian?air?samples[J].J.Chromatogr.A，1999，846：295-306.

Summary of the invention:

The object of the present invention is to provide the detection method of musk xylene in a kind of quick, accurate, highly sensitive cigarette additive.

The present invention adopts oscillation extraction-gas chromatography-mass spectrography technology, uses D ₁₅-musk xylene has been realized the accurate mensuration to musk xylene in the cigarette additive as internal standard compound.

The detection method of musk xylene in a kind of cigarette additive of the present invention, its concrete steps are following:

A. accurately take by weighing 1.0g (being accurate to 0.001g) cigarette additive in the tool plug conical flask of 50mL, add 10mL ultrapure water (resistivity is 18.2M Ω) earlier to its dispersion, add chromatographically pure cyclohexane 10mL then, adding concentration at last is the internal standard compound D of 1.0 μ g/mL ₁₅-musk xylene 0.3mL, stopper seals it beyond the Great Wall;

B. the conical flask with good seal places constant temperature oscillator, and at 25 ℃ of following oscillation extraction 10min, mixed liquor takes out upper organic phase with hydro-extractor centrifugal 10min under the rotating speed of 4000r/min;

C. the aqueous phase after taking out upper organic phase adds chromatographically pure cyclohexane 10mL again, and carries out the extraction second time, centrifugal by the operating conditions of b step, takes out upper organic phase;

D. merge extracted organic phase twice, use to concentrate constant volume behind the anhydrous sodium sulfate drying and be 1mL, be equipped with GC-MS and measure;

E. GC conditions is: carrier gas is high-purity He, flow 1.0mL/min, sample size 1 μ L, not split sampling; Heating schedule: 60 ℃ of initial temperatures, keep 1min, rise to 280 ℃ with 15 ℃/min, keep 10min;

F. the mass spectrum condition is: 280 ℃ of transmission line temperature; Select ion scan, scan feature ion between 11.1～13.5min: quota ion 282,294, auxiliary quota ion 297,312, all the other times are solvent delay; Ion source temperature: 230 ℃; Electron energy 70eV.

The invention has the advantages that: assay method is accurate, quick, highly sensitive, can effectively measure the content of musk xylene in the cigarette additive, is applicable to that tobacco business checks oneself work to the high monitoring of paying close attention to the material musk xylene in the cigarette additive.

Embodiment:

Make further detailed description in the face of the present invention down:

One, experimental section

1.1 instrument and reagent

Instrument and equipment: PE-Clarus600T gas chromatograph-mass spectrometer (U.S. PE), HP-5MS (chromatographic column (U.S. Agilent) of 0.25 μ m * 250 μ m * 30m), XP504 analytical balance (Switzerland's plum Teller); SHZ-82 constant temperature oscillator (Changzhou state China); SK5200H ultrasonic oscillator (Shanghai section leads), LG10-2.4A supercentrifuge (Beijing is medical), separating funnel; Transfer pipet, other common experiment glasswares.

Reagent: cyclohexane, toluene, methenyl choloride, D ₁₅-musk xylene standard items, musk xylene standard items, anhydrous sodium sulfate, ultrapure water (resistivity is 18.2M Ω).

1.2 the preparation of standard solution

Take by weighing musk xylene standard items 0.1g (being accurate to 0.001g) in the 100mL volumetric flask, with cyclohexane dissolving and be settled to scale, be mixed with the musk xylene standard reserving solution of 1mg/mL, in 4 ℃ of preservations.Accurately measure D ₁₅In-musk xylene standard solution (mark article concentration is 100 μ g/mL) 1mL to the 100mL volumetric flask, be settled to scale, be mixed with the D of 1 μ g/mL with cyclohexane ₁₅-musk xylene standard solution.During use; Get 1mg/mL musk xylene standard reserving solution 0.1mL in the 100mL volumetric flask; Be settled to scale with cyclohexane; Be configured to the musk xylene standard solution mother liquor of 1 μ g/mL, get mother liquor 0.2,0.5,1.0,2.0,5.0mL more respectively in the 10mL volumetric flask, adding concentration is the D of 100 μ g/mL ₁₅-musk xylene standard solution 30 μ L are settled to scale with cyclohexane, and the musk xylene concentration of standard solution is respectively 0.02,0.05,0.1,0.2,0.5 μ g/mL; 1.0 the standard solution preparation steps of μ g/mL is: accurately measure the D that concentration is 100 μ g/mL ₁₅-musk xylene standard solution 30 μ L are settled to scale with 1.0 μ g/mL musk xylene mother liquors in the 10mL volumetric flask; Internal standard compound D in the standard solution of above six concentration ₁₅-musk xylene concentration is 0.3 μ g/mL.Form the series standard working solution with these six musk xylene standard solution of above-mentioned 0.02,0.05,0.1,0.2,0.5,1.0 μ g/mL, existing with join at present.

1.3 experimental procedure

Feed liquid/medicinal extract class: take by weighing 1.0g (being accurate to 0.001g) cigarette additive in 50mL tool plug conical flask, after adding ultrapure water (resistivity is 18.2M Ω) the 10mL dispersed sample, adding concentration is the D of 1 μ g/mL ₁₅-musk xylene inner mark solution 0.3mL, cyclohexane 10mL; 10min is extracted in 25 ℃ of vibrations; Mixed liquor takes out upper organic phase with hydro-extractor centrifugal 10min under the rotating speed of 4000r/min, and the aqueous phase after taking out upper organic phase adds chromatographically pure cyclohexane 10mL again; And carry out second time extraction, centrifugal by the operating conditions of above-mentioned steps, take out upper organic phase; Merge extracted organic phase twice, use to concentrate constant volume behind the anhydrous sodium sulfate drying and be 1mL, be equipped with GC-MS and measure.

The tincture class: take by weighing 1.0g (being accurate to 0.001g) cigarette additive in 50mL tool plug conical flask, adding concentration is the D of 1 μ g/mL ₁₅-musk xylene inner mark solution 0.3mL, cyclohexane 10mL, the same feed liquid of all the other experimental procedures/medicinal extract class.

Essential oil/absolute oil class: dilution back direct injected.

GC conditions is: carrier gas is high-purity He, flow 1.0mL/min, sample size 1 μ L, not split sampling; Heating schedule: 60 ℃ of initial temperatures, keep 1min, rise to 280 ℃ with 15 ℃/min, keep 10min;

The mass spectrum condition is: 280 ℃ of transmission line temperature; Select ion scan, scan feature ion between 11.1～13.5min: quota ion 282,294, auxiliary quota ion 297,312, all the other times are solvent delay; Ion source temperature: 230 ℃; Electron energy 70eV.

Two, result and discussion

2.1 the selection of extraction solvent

Be the influence of the extraction solvent of investigating cyclohexane, toluene, 3 kinds of opposed polarities of methenyl choloride to percentage extraction; Adopt 1.3 pre-treating methods; In the cigarette additive sample, add 1 μ g/mL musk xylene standard solution 0.5mL, extract with above-mentioned 3 kinds of solvents respectively, this sample does not add the musk xylene standard solution to be handled as blank with cyclohexane together; Each sample is done twice parallel laboratory test, and the result that makes even all is as testing result.Test findings is seen table 1.

1 extraction solvent is to the influence of percentage extraction

Find out that from table 1 recovery of extraction of cyclohexane and toluene P-xylene Moschus is higher.

2.2 the selection of extraction mode

Investigated vibration, ultrasonic and three kinds of different extraction modes of liquid liquid, two kinds of solvents of cyclohexane and toluene are to the influence of percentage extraction, and as testing result, test findings is seen table 2. to sample-pretreating method with 2.1, twice parallel experiment average results

Table 2 extraction mode is to the influence of percentage extraction

Find out that from table 2 adopt cyclohexane to make solvent, the oscillation extraction effect of extracting is better.

2.3 the selection of extraction time, volume and temperature

Select L for use ₉(3 ⁴) orthogonal test table, adopt cyclohexane to make solvent, investigate the influence of extraction time, temperature and 3 factors of extract volume of oscillation extraction to extraction efficiency, pre-treating method is with 2.1, and test findings is seen table 3.

Table 3 extraction time, extract volume and extraction temperature are to the influence of extraction efficiency

Find out that from table 3 through data analysis, obtaining more excellent scheme is C ₃B ₃A ₁, i.e. the extraction time is 10min, and extraction temperature is 25 ℃, and the extract volume is 20mL.

2.4 the range of linearity and quantitative limit

Adopt deuterium for standard items (D ₁₅-musk xylene) as internal standard compound, musk xylene is with inner mark method ration.Prepare standard operation solution respectively, adopt above chromatographic condition and mass spectrum condition to measure, in standard items concentration was 0.02～1 μ g/mL scope, the linear equation of musk xylene was: Y=1.165X-6.754, r ²=0.999971.With least concentration standard operation solution as unknown sample; Parallel replication 10 times; Concentration value by recording counts standard deviation; Detectability (LOD) is tried to achieve detecting of this detection method P-xylene Moschus with 3 times of standard deviations, quantitative limit (LOQ) with 10 times of standard deviations and is limited to 0.04 μ g/g, quantitatively is limited to 0.13 μ g/g.

2.5 the recovery of method

Adopt cigarette with feed liquid class additive samples, add the test of the recovery and precision, sample adds the standard solution of variable concentrations, makes an experiment by this method, measures with the gas chromatography-mass spectrography sample introduction.The average recovery rate (each adds concentration replicate determination 6 times) that adds in the horizontal extent three of 0.1,0.3,0.5 μ g/g is 84.1%～91.4%, and its recovery result sees table 4.Relative standard deviation RSD value is all less than 10%.

The recovery of standard addition of table 4 musk xylene and relative standard deviation (n=6)

2.6 sample analysis result

Use this method to feed liquid class, medicinal extract class, essential oil/absolute oil class and tincture type tobacco additive agent sample totally 35 carried out analyzing and testing.The result shows that given the test agent does not all detect musk xylene.Adopt same quadrat method, 2 different brands essential wind oil are carried out analyzing and testing, all detect the higher musk xylene of content.

Claims (1)

1. the detection method of musk xylene in the cigarette additive is characterized in that the concrete steps of this method are following:

A. accurately take by weighing the 1.0g cigarette additive in the tool plug conical flask of 50mL, elder generation's adding resistivity is that the ultrapure water 10mL of 18.2M Ω disperses it, adds chromatographically pure cyclohexane 10mL then, and adding concentration at last is the internal standard compound D of 1.0 μ g/mL ₁₅-musk xylene 0.3mL, stopper seals it beyond the Great Wall;

B. the conical flask with good seal places constant temperature oscillator, and at 25 ℃ of following oscillation extraction 10min, mixed liquor takes out upper organic phase with hydro-extractor centrifugal 10min under the rotating speed of 4000r/min;

C. the aqueous phase after taking out upper organic phase adds chromatographically pure cyclohexane 10mL again, and carries out the extraction second time, centrifugal by the operating conditions of b step, takes out upper organic phase;

D. merge extracted organic phase twice, use to concentrate constant volume behind the anhydrous sodium sulfate drying and be 1mL, be equipped with GC-MS and measure;

E. GC conditions is: carrier gas is high-purity He, flow 1.0mL/min, sample size 1 μ L, not split sampling; Heating schedule: 60 ℃ of initial temperatures, keep 1min, rise to 280 ℃ with 15 ℃/min, keep 10min;

F. the mass spectrum condition is: 280 ℃ of transmission line temperature; Select ion scan, scan feature ion between 11.1～13.5min: quota ion 282,294, auxiliary quota ion 297,312, all the other times are solvent delay; Ion source temperature: 230 ℃; Electron energy 70eV.