CN108693291A - A method of measuring tobacco and tobacco product arsenic and chromium different shape compounds content - Google Patents
A method of measuring tobacco and tobacco product arsenic and chromium different shape compounds content Download PDFInfo
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- CN108693291A CN108693291A CN201810778834.3A CN201810778834A CN108693291A CN 108693291 A CN108693291 A CN 108693291A CN 201810778834 A CN201810778834 A CN 201810778834A CN 108693291 A CN108693291 A CN 108693291A
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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
- G01N30/02—Column chromatography
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Abstract
The present invention relates to a kind of measurement tobacco and the methods of tobacco product arsenic and chromium different shape compounds content, belong to heavy metal component determination techniques field in tobacco and tobacco product.The method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content of the present invention, includes the following steps:Sample to be tested is extracted, the pH for then adjusting extracting solution is 6.0~6.8, obtains prepare liquid;Prepare liquid is taken to be analyzed using tablets by HPLC-MS;Efficient liquid phase chromatographic analysis uses gradient elution.The method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content of the present invention, pass through a sample treatment, separation detection simultaneously, analysis method, which have the characteristics that highly sensitive, high-throughput, fast and effective, to be realized to arsenic, 6 kinds of the element different shape substance of chromium two.
Description
Technical field
The present invention relates to a kind of measurement tobacco and the methods of tobacco product arsenic and chromium different shape compounds content, belong to
Heavy metal component determination techniques field in tobacco and tobacco product.
Background technology
Arsenic, chromium have different existing forms in nature, and wherein chromium is mainly with trivalent chromium Cr (III) and Cr VI
The form of Cr (VI) exists, and toxicity is related with form, and Cr (III) is micro- necessary to body metabolism, oral acute toxicity
LD50=1870mg/kg;And Cr (VI) is that absorptivity is high in human body, easy penetration cell film, has genetoxic and carcinogenic work
With oral acute toxicity LD50=190mg/kg sucks carcinogenesis:IUR (sucking unit is dangerous)=12mg/m3.Usual people
The harm of described heavy metal chromium refers to harm of the Cr VI to human body.Arsenic can be divided into organo-arsenic and inorganic arsenic, As (III), As
(V), monomethyl arsenic, dimethyl arsenic and dimethyl arsenic belong to inorganic arsenic, and different its toxicity of arsenic morphology compound also differ.
Since chromium and arsenic that may be present can be directly entered human body as the effect of human saliva is leached in tobacco and tobacco product,
To do harm to huamn body.Therefore, while realization just seems particularly necessary to the measurement of tobacco and tobacco product arsenic and chromium.
The analysis about arsenic, chromium form has a large amount of document report, but arsenic, chromium shape at present in the prior art
The document of state analysis all concentrates on food, marine product and environmental area, and the Research Literature in tobacco and buccal cigarette field is gone back at present
It is fewer, as application publication number be CN104833750A Chinese invention patent application disclose one grow tobacco and tobacco product in
The method of separating and assaying of different valence state chromium, this method use High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument pair
The content of trivalent chromium and Cr VI is measured in sample, and the mobile phase that when high performance liquid chromatography separation uses is 7.0 for pH
The ammonium nitrate solution of 0.1mol/L, flow velocity 1.0mL/min, since the concentration of mobile phase is excessive, eluent ionic strength mistake
Greatly so that the compounds such as monomethyl arsenic and dimethyl arsenic can hardly retain on ion chromatographic column, cause to can not achieve same
Simultaneously to the separation determination of arsenic and chromium different shape compound under the conditions of one.
Invention content
The object of the present invention is to provide a kind of measurement tobacco and tobacco product arsenic and chromium different shape compounds contents
Method can be carried out at the same time measurement to arsenic and chromium different shape compounds content.
In order to achieve the goal above, the technical solution adopted in the present invention is:
A method of tobacco and tobacco product arsenic and chromium different shape compounds content are measured, is included the following steps:
Sample to be tested is extracted, the pH for then adjusting extracting solution is 6.0~6.8, obtains prepare liquid;Prepare liquid is taken to adopt
It is analyzed with tablets by HPLC-MS;
In analytic process, efficient liquid phase chromatographic analysis uses gradient elution;The mobile phase A that the gradient elution uses is pH
=6.0~6.8, the ammonium nitrate solution of a concentration of 5~15mmol/L, Mobile phase B be pH=6.0~6.8, it is a concentration of 50~
The ammonium nitrate solution of 75mmol/L;Gradient elution program includes:0min~C min mobile phases are 100%A, C min~D
Min is 100%B;C min are selected from any time node between 3.5min~4.5min;Dmin is selected from 18~25min
Between any time node;The flow velocity of mobile phase is 0.5~0.8mL/min.
The method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content of the present invention, passes through a sample
Product processing, to arsenic, chromium two 6 kinds of element different shape substance (trivalent arsenic, monomethyl arsenic, dimethyl arsenic, pentavalent arsenic, trivalent chromium
And Cr VI) realize that separation detection simultaneously, analysis method have the spies such as highly sensitive, high-throughput, fast and effective, applied widely
Point.
Mobile phase A and Mobile phase B are obtained after being adjusted to the pH of the ammonium nitrate solution of respective concentration.To flowing
Phase pH is adjusted using ammonium hydroxide or nitric acid.
The tablets by HPLC-MS uses High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument.
Inductive coupling plasma mass spectrometry condition is:Transmission power is 1320kW;Carrier gas is Ar, flow rate of carrier gas is 1.0~
1.1mL/min;Collision gas flow velocity is 4.5mL/min, and collision gas is helium;Wriggling revolution speed is 0.3~0.4rps;Analyze matter
Measuring number is75As,52Cr;The time of integration uses75As and52Cr is 0.5min;Sampling time 30min.
The method that sample to be tested extracts is included the following steps:It is 0.2~0.4% by sample to be tested and mass fraction
Mixed in hydrochloric acid after be ultrasonically treated 20~50min.
Every 0.5~1.0g samples to be tested correspond to the volume of the hydrochloric acid used as 20~30mL.
The chromatographic column that efficient liquid phase chromatographic analysis uses is the agilent biowax chromatographic column of 4 × 25cm.Efficient liquid phase
Sample size when chromatography is analyzed is 10~100 μ L.It is further preferred that sample size of high performance liquid chromatography when sample introduction is analyzed
For 10~50 μ L.
It is Cr (III) complex compound, six that tablets by HPLC-MS, which carries out analysis accepted standard work liquor,
The mixed standard solution of valence chromium, trivalent arsenic, monomethyl arsenic, dimethyl arsenic, pentavalent arsenic;Cr (III) complex compound uses carboxylic acid network
Mixture obtains the complexing completely of trivalent chromium standard items.
The carboxylic acid complexing agent is ethylenediamine tetra-acetic acid (EDTA).
The method of said determination tobacco and tobacco product arsenic and chromium different shape compounds content, further includes taking prepare liquid
Before being analyzed using tablets by HPLC-MS, prepare liquid is used into aqueous membrane filtration;The aqueous filter membrane
Aperture is 0.45 μm.
The method of said determination tobacco and tobacco product arsenic and chromium different shape compounds content further includes marking series
Quasi- solution is analyzed using HPLC MS, is respectively then vertical with series standard solution concentration and peak area
Abscissa draws standard curve.The condition and use high that series standard solution is analyzed using HPLC MS
The condition that effect liquid phase chromatogram-mass spectrography analyzes prepare liquid is identical.With trivalent arsenic, monomethyl arsenic, dimethyl in prepare liquid
The peak area size of arsenic, pentavalent arsenic and trivalent chromium, Cr VI on instrument, establishing criteria curve are quantified.
Description of the drawings
Fig. 1 is the extraction ion of trivalent arsenic, monomethyl arsenic, dimethyl arsenic, pentavalent arsenic in mixed standard solution in embodiment 1
(As75) chromatogram;
Fig. 2 is the extraction ion (Cr of trivalent chromium, Cr VI in mixed standard solution in embodiment 152) chromatogram.
Specific implementation mode
Technical scheme of the present invention is further described below in conjunction with the mode of specific implementation.
The chromatographic column in High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument employed in specific implementation mode is
agilent biowax(4×25cm);When analysis, inductivity coupled plasma mass spectrometry condition is:Transmission power:1320kW;It carries
Gas velocity:1.10mL/min(Ar);Collision gas flow velocity:4.5mL/min(He);Wriggling revolution speed:0.3rps;The time of integration:
0.5min;Sampling time:15min;Analyze mass number:75As,52Cr;
Mixed standard solution employed in each embodiment is prepared using the method included the following steps:
I) the trivalent chromium standard items solution soluble in water for preparing a concentration of 100 μ g/L of trivalent chromic ion is taken, is subsequently placed in 80
DEG C water bath with thermostatic control in, be warming up to after 80 DEG C be added 2mmol/L EDTA solution reactions 40min so that trivalent chromium is fully complexed, obtain
To Cr (III) complex solution;
Ii certain density trivalent arsenic standard solution, monomethyl arsenic standard solution, dimethyl arsenic standard) are prepared respectively
Then product solution, pentavalent arsenic standard solution and Cr VI standard solution take suitable Cr (III) complex solution, three respectively
Valence arsenic standard solution, monomethyl arsenic standard solution, dimethyl arsenic standard solution, pentavalent arsenic standard solution and Cr VI
Standard solution mixes, and obtains mixed standard solution;The mixed standard solution of gained is diluted, is obtained with a certain concentration
The series standard working solution of gradient, each standard working solution concentration gradient are shown in Table 1.
1 each standard working solution concentration gradient (unit of table:μg/L)
Mobile phase A and Mobile phase B in each embodiment are that respective concentration first is made by ammonium nitrate is soluble in water when preparing
Ammonium nitrate solution, be then respectively adopted ammonium hydroxide or nitric acid adjustment pH to corresponding range in.
Embodiment 1
The method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content of the present embodiment, is in bulk
The measurement of arsenic and chromium different shape compounds content, includes the following steps in buccal cigarette:
1) it weighs the buccal cigarette sample in bulk after 0.5g grindings to be put into conical flask, the salt that mass fraction is 0.2% is added
Sour 25mL is put into ultrasonic extraction 30min in ultrasonic wave after flooding offal sample completely, is separated by solid-liquid separation, obtains extract liquor;It is super
Acoustic power is 300W, frequency 40KHz;
2) extract liquor obtained by step 1) is crossed into 0.45 μm of aqueous filter membrane, obtains filtrate, then adjust filtrate with ammonium hydroxide
Between pH to 6.0~6.8, filtrate is placed in as prepare liquid in chromatogram bottle after taking 1mL to adjust pH;
3) take the prepare liquid 1mL of institute in step 2) using the progress of High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument
Analysis;
Chromatographic condition is used by analysis:Mobile phase A is 15mmol/L ammonium nitrate (pH 6.5), and Mobile phase B is
50mmol/L ammonium nitrate (pH 6.5);Flow velocity:0.6mL/min;Eluent gradient program is t=0.0min, 100%A, t=
4.0min, 100%B, t=25min, 100%A, t=30min, 100%A;Column oven:Room temperature, sample size:50μL;
4) condition analyzed using High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument in step 3)
Under, with a concentration of abscissa, peak area is ordinate, draws standard curve;Trivalent arsenic in mixed standard solution, monomethyl arsenic,
Extraction ion (the As of dimethyl arsenic, pentavalent arsenic75) chromatogram is shown in Fig. 1;Trivalent chromium in mixed standard solution, Cr VI extraction from
Son (Cr52) chromatogram is shown in Fig. 2;
It is quantified with the peak area size measured in step 3), analysis result is shown in Table 2.
The measurement result of table 2 buccal cigarette arsenic in sample in bulk and chromium element
Embodiment 2
The method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content of the present embodiment, is to tobacco
The measurement of middle arsenic and chromium different shape compounds content, includes the following steps:
1) it weighs the tobacco sample after 0.5g grindings to be put into conical flask, a concentration of 0.2% hydrochloric acid 25mL is added, waits for molten
After liquid is totally submerged sample, it is put into progress ultrasonic extraction 30min in ultrasonic wave, is separated by solid-liquid separation, obtains extract liquor;Ultrasonic power
300W, frequency 40KHz;
2) extract liquor obtained by step 1) is crossed into 0.45 μm of water phase filter membrane, obtains filtrate, then adjust filtrate with ammonium hydroxide
Between pH to 6.0~6.8, filtrate is placed in as prepare liquid in chromatogram bottle after taking 1mL to adjust pH;
3) prepare liquid is taken to be analyzed using High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument;Analysis is used
Chromatographic condition be:Mobile phase A is 15mmol/L ammonium nitrate (pH 6.5), and Mobile phase B is that (pH is 50mmol/L ammonium nitrate
6.5);Flow velocity:0.6mL/min;Eluent gradient program is t=0.0min, 100%A, t=4.0min, 100%B, t=
25min, 100%A, t=30min, 100%A;Column oven:Room temperature, sample size:50μL;
4) condition analyzed using High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument in step 3)
Under, with a concentration of abscissa, peak area is ordinate, draws standard curve;It is carried out with the peak area size measured in step 3)
Quantitative, analysis result is shown in Table 3.
The measurement result of arsenic and chromium in 3 tobacco of table
| Determinand | Measured value (μ g/kg) |
| As(Ⅲ) | 11.5 |
| As(Ⅴ) | 40.8 |
| Monomethyl arsenic | 7.8 |
| Dimethyl arsenic | 10.4 |
| Cr(Ⅲ) | 36.4 |
| Cr(Ⅵ) | 2.7 |
According to the method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content in embodiment 2, with reality
Repeatability and the rate of recovery that sample in example 2 has investigated method are applied, sample introduction is repeated 10 times with standard curve minimum point, with 10 sample introductions
As a result detection limit of the standard deviation as method, repeatability, the rate of recovery and detection limit the results are shown in Table 4.
4 repeatability of table, the rate of recovery and detection limit
Note:The coefficient of variation and rate of recovery unit are % in table 4, and detection limit unit is μ g/L, other numerical value units are μ g/
kg。
Embodiment 3
The method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content of the present embodiment, including it is following
Step:
1) 1g buccal cigarette samples are weighed to be put into conical flask, a concentration of 0.4% hydrochloric acid 50mL is added, waits for that solution soaks completely
After not having sample, it is put into progress ultrasonic extraction 30min in ultrasonic wave, is separated by solid-liquid separation, obtains extract liquor;Ultrasonic power 300W, frequency
Rate 40KHz;
2) extract liquor obtained by step 1) is crossed into 0.22 μm of water phase filter membrane, obtains filtrate, then adjust filtrate with ammonium hydroxide
Between pH to 6.0~6.8, filtrate is placed in as prepare liquid in chromatogram bottle after taking 1mL to adjust pH;
3) prepare liquid is taken to be analyzed using High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument;Analysis is used
Chromatographic condition be:Mobile phase A is 10mmol/L ammonium nitrate (pH6.8), and Mobile phase B is 60mmol/L ammonium nitrate (pH6.0);Stream
Speed:0.6mL/min;Eluent gradient program is t=0.0min, 100%A, t=4.0min, 100%B, t=20min, 100%
A, t=30min, 100%A;Column oven:Room temperature, sample size:10μL;
4) condition analyzed using High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument in step 3)
Under, with a concentration of abscissa, peak area is ordinate, draws standard curve;It is carried out with the peak area size measured in step 3)
It is quantitative.
Embodiment 4
The method for measuring tobacco and tobacco product arsenic and chromium different shape compounds content of the present embodiment, including it is following
Step:
1) 1.0g tobacco samples are weighed to be put into conical flask, a concentration of 0.4% hydrochloric acid 50mL is added, waits for that solution soaks completely
After not having sample, it is put into progress ultrasonic extraction 30min in ultrasonic wave, is separated by solid-liquid separation, obtains extract liquor;Ultrasonic power 300W, frequency
Rate 40KHz;
2) extract liquor obtained by step 1) is crossed into 0.45 μm of water phase filter membrane, obtains filtrate, then adjust filtrate with ammonium hydroxide
Between pH to 6.0~6.8, filtrate is placed in as prepare liquid in chromatogram bottle after taking 1mL to adjust pH;
3) prepare liquid is taken to be analyzed using High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument;Analysis is used
Chromatographic condition be:Mobile phase A is 5mmol/L ammonium nitrate (pH6.0), and Mobile phase B is 75mmol/L ammonium nitrate (pH6.8);Stream
Speed:0.6mL/min;Eluent gradient program is t=0.0min, 100%A, t=3.5min, 100%B, t=18min, 100%
A, t=30min, 100%A;Column oven:Room temperature, sample size:10μL;
4) condition analyzed using High performance liquid chromatography-inductively coupled plasma mass spectrometry instrument in step 3)
Under, with a concentration of abscissa, peak area is ordinate, draws standard curve;It is carried out with the peak area size measured in step 3)
It is quantitative.
Claims (8)
1. a kind of method measuring tobacco and tobacco product arsenic and chromium different shape compounds content, it is characterised in that:Including
Following steps:
Sample to be tested is extracted, the pH for then adjusting extracting solution is 6.0~6.8, obtains prepare liquid;Take prepare liquid using high
Effect liquid phase chromatogram-Mass Spectrometry is analyzed;
In analytic process, efficient liquid phase chromatographic analysis uses gradient elution;
The mobile phase A that the gradient elution uses is pH=6.0~6.8, the ammonium nitrate solution of a concentration of 5~15mmol/L, stream
Phase B is moved as pH=6.0~6.8, the ammonium nitrate solution of a concentration of 50~75mmol/L;
Gradient elution program includes:0min~C min mobile phases are 100%A, and C min~D min are 100%B;C
Min is selected from any time node between 3.5min~4.5min;Dmin is selected from any time between 18~25min
Point;The flow velocity of mobile phase is 0.5~0.8mL/min.
2. the method according to claim 1 for measuring tobacco and tobacco product arsenic and chromium different shape compounds content,
It is characterized in that:The tablets by HPLC-MS uses High performance liquid chromatography-inductively coupled plasma mass spectrometry
Instrument.
3. the method according to claim 2 for measuring tobacco and tobacco product arsenic and chromium different shape compounds content,
It is characterized in that:Inductive coupling plasma mass spectrometry condition is:Transmission power is 1320kW;Carrier gas is Ar, flow rate of carrier gas is 1.0~
1.1mL/min;Collision gas flow velocity is 4.5mL/min, and collision gas is helium;Wriggling revolution speed is 0.3~0.4rps;Analyze matter
Measuring number is75As,52Cr;The time of integration uses75As and52Cr is 0.5min;Sampling time 30min.
4. the method according to claim 1 for measuring tobacco and tobacco product arsenic and chromium different shape compounds content,
It is characterized in that:The method that sample to be tested extracts is included the following steps:By sample to be tested and mass fraction be 0.2~
20~50min is ultrasonically treated after 0.4% mixed in hydrochloric acid.
5. the method according to claim 4 for measuring tobacco and tobacco product arsenic and chromium different shape compounds content,
It is characterized in that:Every 0.5~1.0g samples to be tested correspond to the volume of the hydrochloric acid used as 20~30mL.
6. the method according to claim 1 for measuring tobacco and tobacco product arsenic and chromium different shape compounds content,
It is characterized in that:The chromatographic column that efficient liquid phase chromatographic analysis uses is the agilent biowax chromatographic column of 4 × 25cm.
7. the method according to claim 1 for measuring tobacco and tobacco product arsenic and chromium different shape compounds content,
It is characterized in that:Tablets by HPLC-MS carry out analysis accepted standard work liquor be Cr (III) complex compound,
The mixed standard solution of Cr VI, trivalent arsenic, monomethyl arsenic, dimethyl arsenic, pentavalent arsenic;Cr (III) complex compound uses carboxylic acid
Complexing agent obtains the complexing completely of trivalent chromium standard items.
8. the method according to claim 1 for measuring tobacco and tobacco product arsenic and chromium different shape compounds content,
It is characterized in that:Further include that prepare liquid is used into water before taking prepare liquid to be analyzed using tablets by HPLC-MS
Property membrane filtration;The aperture of the aqueous filter membrane is 0.45 μm.
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| CN108693291B (en) | 2020-07-17 |
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