CN106908303A - A kind of pre-treating method of milk sample is Ji the assay method of bioxin - Google Patents
A kind of pre-treating method of milk sample is Ji the assay method of bioxin Download PDFInfo
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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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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- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
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- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
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Abstract
The invention provides a kind of pre-treating method of milk sample Ji the assay method of bioxin.The pre-treating method of the milk sample, comprises the following steps：Sample extraction, sample purification, nitrogen blow concentration；In sample extraction process, milk sample is hydrolyzed, efficiently extracts out protein component, fat constituent, with quick, efficient, chemo-selective it is high, extraction efficiency is high the features such as；And it is simple and easy to apply, solvent consumption can be reduced, cost is reduced, with high application value.
Description
Technical field
The present invention relates to pollutant monitoring technical field, and in particular to a kind of pre-treating method of milk sample is Ji bioxin
Assay method.
Background technology
Many chlorodiphenyls and Dui bioxin and polychlorinated dibenzo (PCDD/Fs) are the oxygen-containing aromatics chemical combination of a class chloro
Thing, due to the difference of chlorine atom number and the position of substitution in molecule, has 210 kinds of congeners, and often detection is 17 kinds in the world
Many chloro dibenzo Dui bioxin and polychlorinated dibenzo of most strong 2,3,7, the 8 chlorine atoms substitution of toxicity, because of its toxicity
High, the World Health Organization (WHO) has been classified as deadly poisonous compound, and specifies that acceptable daily intake is 1~40pg/
Kg body weight.This kind of material is highly stable, and fusing point is higher, it is extremely difficult to be dissolved in water, is the liposoluble substance of colorless and odorless, easily in biology
Cylinder accumulation, with human health risk higher.Environmental Protection Agency's research shows that mankind bioxin exposure 90% is to pass through
Animal food intake.
Contain the extracting methods such as substantial amounts of protein and fat constituent, common soxhlet type in milk sample, it is impossible to have
Effect ground aminosal, isolates fat constituent, therefore, it is necessary to provide a kind of pre-treating method of milk sample Ji bioxin
Assay method, to reduce influence of the impurity such as protein and fat to testing result.
The content of the invention
To solve the problems, such as prior art, the invention provides a kind of pre-treating method of milk sample Ji bioxin
Assay method.
To achieve these goals, present invention employs following technical scheme：
In a first aspect, the invention provides a kind of pre-treating method of milk sample, comprising the following steps：
(1) after milk sample is hydrolyzed, the first solvent extraction is added, isolates the first water phase and the first organic phase, then to
The second solvent extraction is added in first water phase, the second water phase and Second Organic Phase is isolated, merging the first organic phase and second has
Machine phase, concentrate is obtained after blowing concentration through nitrogen, the concentrate is well mixed with mix reagent, centrifugation, removes dense sulphur
Acid layer, is obtained extract solution；Wherein, first organic solvent and second organic solvent are identical or different organic molten
Agent；The organic solvent includes but is not limited to the one kind in ether, acetone, dimethyl sulfoxide (DMSO), toluene, n-hexane, dichloromethane
Or it is several；
(2) extract solution obtained by step (1) is carried out into acidic silica gel post, acidic alumina silica gel column purification successively, is obtained
Scavenging solution；
(3) scavenging solution obtained by step (2) is blown into concentration through nitrogen, is added13C-PCDD/Fs internal standards, obtain Instrumental Analysis use
Purification sample.
Preferably, in the step (1), the milk sample is commercially available liquid milk or natural liquid milk.
Preferably, in the step (1), the step of the hydrolysis, specifically include：
Milk sample is mixed with alkaline protease solution, 3 are hydrolyzed under conditions of 40~55 DEG C, pH=8.5~10.0
~5h；Add alkaline solution and hydrolyze 15~35min at 70~80 DEG C, add Organic Alcohol concussion and be well mixed, wherein, it is described
The mass fraction of alkaline protease solution neutral and alkali protease is 1~5% (more preferably 2%)；The alkali protease
Solution is 1 with the volume ratio of the milk sample:10~20；The alkaline solution includes but is not limited to ammonia spirit, hydroxide
One or more in sodium water solution, potassium hydroxide aqueous solution；The Organic Alcohol include but is not limited to ethanol, methyl alcohol, butanediol,
One or more in propyl alcohol.
It is further preferred that in the step (1), the alkaline solution is ammonia spirit, wherein, the quality of ammonia spirit
Percentage concentration is 25~30%.
It is further preferred that in the step (1), the volume ratio of the alkaline solution and the milk sample for 0.1~
2:1 (is still more preferably 0.5:1).
It is further preferred that in the step (1), the Organic Alcohol is ethanol.
It is further preferred that in the step (1), the Organic Alcohol is 0.5~4 with the volume ratio of the milk sample:1
(more preferably 1:1).
Preferably, in the step (1), the described organic solvent of addition first extraction separates water outlet phase and first organic
The step of phase, specifically include：
The first organic solvent, 3~10min of oscillation extraction (preferably 5min) is added to separate water outlet phase and the first organic phase.
Preferably, in the step (1), first organic solvent is molten for the mixing of ether, n-hexane and dichloromethane
Agent, wherein, the volume ratio of ether, n-hexane and dichloromethane is 1~3:1~3:1 (is still more preferably 2:2:1).
Preferably, in the step (1), to adding the second organic solvent to extract in the first water phase, the second water phase is isolated
And the step of Second Organic Phase, specifically include：
Second is isolated to the second organic solvent, 3~10min of oscillation extraction (preferably 5min) is added in the first water phase
Water phase and Second Organic Phase.
Preferably, in the step (1), second organic solvent is the mixed solvent of n-hexane and dichloromethane, its
In, the volume ratio of n-hexane and dichloromethane is 0.5~3:1 (is still more preferably 1:1).
It is understood that the present invention is not particularly limited to the method for the centrifugation described in step (1), using this
The conventional centrifugal separation method of art personnel.
Preferably, in the step (1), described is well mixed the concentrate with mix reagent, and centrifugation is gone
Except the concentrated sulfuric acid layer, be obtained extract solution the step of, specifically include：
By the concentrate and solvent, inorganic acid solution vortex shaken well, then centrifuge is used with 2000~4000r/
The centrifugation rate of min (more preferably 3000r/min) is centrifuged 0.5~4min (more preferably 1min), then with drop
Pipe sucks the concentrated sulfuric acid of lower floor, be obtained extract solution, wherein, the solvent include but is not limited to ether, acetone, dimethyl sulfoxide (DMSO),
One or more in toluene, n-hexane, dichloromethane；The solvent is 1~10 with the volume ratio of the concentrate:1；It is described
Inorganic acid solution is 1~10 with the volume ratio of the concentrate:1.
It is further preferred that in the step (1), the solvent is n-hexane.
It is further preferred that in the step (1), the inorganic acid solution is concentrated sulfuric acid solution, wherein, concentrated sulfuric acid solution
Middle concentrated sulfuric acid mass fraction is 70~98%.
Preferably, the step (2), specifically includes：
The extract solution obtained by step (1) is carried out only using the acidic silica gel post after being washed with the first pre- eluent pre-leaching
Change, through the first eluent drip washing after, be concentrated to give the first scavenging solution；The scavenging solution is transferred to using the second eluent pre-
In the acidic alumina silicagel column of drip washing treatment, after carrying out drip washing using the second eluent, by the acidic alumina silicagel column
After in turn, using the 3rd eluent drip washing, and collection obtains the second scavenging solution；
Wherein, the described first pre- eluent include but is not limited to one kind in ether, toluene, n-hexane and dichloromethane or
It is various；First eluent is included but is not limited to one or more in ether, toluene, n-hexane and dichloromethane；It is described
Second eluent includes but is not limited to one or more in n-hexane, dichloromethane, methyl alcohol, toluene；3rd eluent
One or more including but not limited in n-hexane, dichloromethane, toluene.
It is further preferred that in the step (2), in the acidic silica gel post height of acidic silica gel be 5~10cm (more
More preferably 8cm).
It is further preferred that in the step (2), the first pre- eluent and the first eluent are identical or different
Organic solvent.
It is further preferred that in the step (2), the first pre- eluent is n-hexane.
It is further preferred that in the step (2), the consumption of the first pre- eluent for 10~25mL (further
Preferably 15mL).
It is further preferred that in the step (2), first eluent is n-hexane.
It is further preferred that in the step (2), the consumption of first eluent is that 20~30mL is (further excellent
Elect 23mL as).
It is further preferred that in the step (2), second eluent is the mixing of dichloromethane, methyl alcohol and toluene
Solvent, wherein, the dichloromethane, methyl alcohol and volume of toluene ratio are 12~16:2~5:1, (still more preferably it is 15:4:
1)。
It is further preferred that in the step (2), the 3rd eluent is toluene.
It is further preferred that in the step (2), the consumption of the 3rd eluent is that 25~50mL is (further excellent
Elect 30mL as).
It is further preferred that in the step (2), the height of acidic oxidation aluminum silica gel in the acidic alumina silicagel column
It is 5~10cm (being still more preferably 8cm).
It is further preferred that in the step (2), the acidic alumina silicagel column of the pre- drip washing treatment is under
Method is stated to be obtained：
Pre- drip washing for the first time is carried out to the acidic alumina silicagel column using the second pre- eluent, then using the 3rd pre-leaching
Lotion carries out second pre- drip washing to the acidic alumina silicagel column, then using the 4th pre- eluent to the acidic alumina
Silicagel column carries out the pre- drip washing of third time, then the 4th pre-leaching is carried out to the acidic alumina silicagel column using the 5th pre- eluent
Wash, wherein, the second pre- eluent, the 3rd pre- eluent, the 4th pre- eluent, the 5th pre- eluent are identical or different
Pre- drip washing organic solvent, wherein, the pre- drip washing organic solvent is included but is not limited in n-hexane, dichloromethane, methyl alcohol, toluene
One or more.
It is further preferred that in the step (2), the second pre- eluent is the mixing of n-hexane and dichloromethane
Solvent, wherein, the volume ratio of the n-hexane and dichloromethane is 14~18:1.
It is further preferred that in the step (2), the consumption of the second pre- eluent is 5~10mL.
It is further preferred that in the step (2), the 3rd pre- eluent is the mixing of n-hexane and dichloromethane
Solvent, wherein, the volume ratio of the n-hexane and dichloromethane is 1~3:1.
It is further preferred that in the step (2), the consumption of the 3rd pre- eluent is 10~20mL.
It is further preferred that in the step (2), the 4th pre- eluent is toluene.
It is further preferred that in the step (2), the consumption of the 4th pre- eluent is 7~15mL.
It is further preferred that in the step (2), the 5th pre- eluent is n-hexane.
It is further preferred that in the step (2), the consumption of the 5th pre- eluent is 5~10mL.
Preferably, the step (3), specifically includes：
Scavenging solution obtained by step (2) is transferred in sample bottle, nitrogen is blown to do, is transferred to the sample that capacity is 1.5mL
In bottle, 10~14h is placed, add internal standard constant volume, the purification sample of prepared Instrumental Analysis.
Second aspect, present invention also offers a kind of assay method of milk sample Zhong bioxin, comprises the following steps：
The obtained purification sample of milk sample pre-treating method as described in relation to the first aspect is taken, using isotopic dilution high score
Distinguish that gas chromatography-high resolution mass spectrometry combined instrument carries out Instrumental Analysis measure to the purification sample, show that Niu milk Zhong bioxin contains
The measurement result of amount.
Preferably, the location parameter of the isotopic dilution high resolution spectral estimation combined instrument is as follows：
GC conditions are：Input mode is the μ L of Splitless injecting samples 1, and injector temperature is 280 DEG C, and carrier gas flux is
1.0mL/min, chromaticness interface temperature is 280 DEG C, and chromatographic column is DB-5ms (60m × 0.25mm × 0.1 μm)；Initial temperature 90
DEG C, 220 DEG C are warming up to the speed of 18 DEG C/min after keeping 2min, then 260 DEG C are warming up to 1.4 DEG C/min, stop 4min；Again
305 DEG C are warming up to 4 DEG C/min, 4min is stopped；
Mass Spectrometry Conditions are：Chromaticness interface temperature is 300 DEG C, and ion source temperature is 280 DEG C, and EI sources collision energy is 70eV,
Using selection ion (SIM) mode to [M]+、[M+2]+[M+4]+Characteristic ion is monitored.
The beneficial effects of the present invention are：
(1) milk sample pre-treating method provided by the present invention, by milk sample using enzymolysis and aminosal into
Point, fat constituent is efficiently extracted out, extraction process has the features such as quick, efficient, chemo-selective is high, extraction efficiency is high；
(2) pre-treating method of milk sample provided by the present invention can subtract Ji the assay method of bioxin, simple and easy to apply
Few solvent consumption, reduces cost, with high application value.
Specific embodiment
The present invention is described in further detail below by specific embodiment, to be clearly understood that the present invention is protected
Technical scheme.
Embodiment 1
The invention provides a kind of pre-treating method of milk sample Ji the assay method of bioxin, comprises the following steps：
(1) extract：The commercially available liquid milks of 200mL are taken, 15mL 2% (w/w) alkali protease is added, in 50 DEG C, pH=
4h is hydrolyzed under conditions of 9.0,100mL ammoniacal liquor (mass fraction is 25%) is added, 70~80 DEG C of hydrolysis 20min add 200mL
Alcohol solvent (in alcohol solvent the mass fraction of ethanol be 95%), vibration 10min is mixed, add 200mL ether, n-hexane and
(in the mixed solvent, the volume ratio of ether, n-hexane and dichloromethane is 2 to the mixed solvent of dichloromethane:2:1), vibrate
5min, takes the first organic phase, adds the mixed solvent (body of n-hexane and dichloromethane of 200mL n-hexanes and dichloromethane
Product is than being 1:1) 5min, is vibrated, Second Organic Phase is taken, and the first organic phase of merging and Second Organic Phase are fitted into sample bottle, then
Concentration is blown through nitrogen closely to do, obtain concentrating sample；Again to addition 7mL n-hexanes and the 8mL concentrated sulfuric acids (wherein, the concentrated sulfuric acid in concentrate
The mass fraction of middle sulfuric acid is 75%), after vortex shaken well, to use centrifuge to be centrifuged with the centrifugation rate of 3000r/min
1min, then sucks the concentrated sulfuric acid (repeating this step to without obvious yellow) of lower floor with dropper, and extract solution is obtained；
(2) purify：
The step of dress post, is as follows：Mineral wool is added, glass bar decurl is used, is cleaned with dichloromethane and is blown with hair-dryer
It is dry, 8cm acid silicon has been measured, the position (carrying out mark with marker) of 8cm acidic aluminas installs sour silicon and acidic oxidation respectively
Aluminium, is obtained acidic silica gel post and acidic alumina silicagel column；
The step of acidic silica gel column purification, is as follows：
By the column jecket of acidic silica gel post and connect liquid bath and coupled together with connector, waste liquid is placed under the acidic silica gel post
Cylinder, using dropper by 2 addition acidic silica gel posts of 15mL n-hexanes point, when the n-hexane quick-drying in post, is replaced with sample bottle
Waste liquid cylinder is changed, then with 3 flushing sample bottles of 21mL n-hexanes point, the extract solution obtained by step (1) is transferred to acidic silica gel post
In, the wall of pillar is then rinsed with 2mL n-hexanes；After being dripped off Deng the solution in acidic silica gel post, collection obtains the first scavenging solution,
And acidic silica gel post is removed；
The step of acidic alumina silica gel column purification, is as follows：
By the column jecket of acidic alumina silicagel column and connect liquid bath and coupled together with connector, in the acidic oxidation aluminum silica gel
Waste liquid cylinder is placed under post, using dropper by the mixed solvent (volume of n-hexane and dichloromethane of 8mL n-hexanes and dichloromethane
Than being 94:6, points 4 times, each 2mL) add acidic alumina column；When the mixing of n-hexane and dichloromethane in post is molten
After agent quick-drying, then (wherein, the volume ratio of n-hexane and dichloromethane is using the mixed solvent of 16mL n-hexanes and dichloromethane
3:2, point 4 each 4mL) rinse pillar；First added for 3-4 times with 10mL toluene point and rinse pillar；Add the flushing of 6mL n-hexanes
Pillar；
Rinsed just pillar in turn, use mixed solvent (n-hexane and the dichloromethane of 8mL n-hexanes and dichloromethane
The volume ratio of alkane is 1:1, point 4 additions, each 2mL) sample bottle is rinsed, first scavenging solution is transferred to acidic alumina
In silicagel column；(wherein, dichloromethane, methyl alcohol and volume of toluene ratio are to add the mixed solvent of dichloromethane, methyl alcohol and toluene
15:4:1)；After solution stream is complete, waste liquid cylinder is replaced with sample flask, and, add 30mL toluene, and use sample pillar in turn
Product flask collects the second scavenging solution；
(3) concentrate：Add 10mL dichloromethane to rinse sample flask, the second scavenging solution obtained by step (2) is transferred to
In sample bottle, nitrogen is blown to do, and adds 1mL dichloromethane, and is transferred in the sample bottle of 1.5mL, places or so evening
Time, after dichloromethane is evaporated completely, with the internal standard constant volume of 20 μ L, the purification sample of prepared Instrumental Analysis；
(4) determine：The purification sample obtained by step (3) is taken, using the gas chromatograph-high resolved matter of isotopic dilution high-resolution
Spectrum combined instrument carries out Instrumental Analysis measure to the purification sample, draws the measurement result of liquid milk Zhong content of dioxin；Wherein,
The location parameter of the isotopic dilution high resolution spectral estimation combined instrument is as follows：
GC conditions are：Input mode is the μ L of Splitless injecting samples 1, and injector temperature is 280 DEG C, and carrier gas flux is
1.0mL/min, chromaticness interface temperature is 280 DEG C, and chromatographic column is DB-5ms (60m × 0.25mm × 0.1 μm)；Initial temperature 90
DEG C, 220 DEG C are warming up to the speed of 18 DEG C/min after keeping 2min, then 260 DEG C are warming up to 1.4 DEG C/min, stop 4min；Again
305 DEG C are warming up to 4 DEG C/min, 4min is stopped；
Mass Spectrometry Conditions are：Chromaticness interface temperature is 300 DEG C, and ion source temperature is 280 DEG C, and EI sources collision energy is 70eV,
Using selection ion (SIM) mode to [M]+、[M+2]+[M+4]+Characteristic ion is monitored.
The assay method of the embodiment of the present invention 1 applies the testing result in milk sample as shown in table 1.Using the present invention
The rate of recovery of preprocess method , bioxin Isotopic Internal Standards is between deposit 78%~94%.
The testing result of the sample Zhong content of dioxin of table 1
The above, is only presently preferred embodiments of the present invention, and any formal limitation is not made to the present invention, though
So the present invention is disclosed above with preferred embodiment, but is not limited to the present invention, any to be familiar with this professional technology people
Member, without departing from the scope of the present invention, when making a little change or modification using the technology contents of the disclosure above
Be the Equivalent embodiments of equivalent variations, as long as be without departing from technical solution of the present invention content, according to technical spirit of the invention,
Within the spirit and principles in the present invention, any simple modification, equivalent and improvement for being made to above example etc., still
Belong within the protection domain of technical solution of the present invention.
Claims (10)
1. a kind of milk sample pre-treating method, it is characterised in that comprise the following steps：
(1) after milk sample is hydrolyzed, the first solvent extraction is added, isolates the first water phase and the first organic phase, then to first
The second solvent extraction is added in water phase, the second water phase and Second Organic Phase is isolated, merges the first organic phase and Second Organic Phase,
Concentrate is obtained after blowing concentration through nitrogen, the concentrate is well mixed with mix reagent, centrifugation, removal concentrated sulfuric acid layer,
Extract solution is obtained；Wherein, first organic solvent and second organic solvent are identical or different organic solvent；It is described
Organic solvent includes one or more in ether, acetone, dimethyl sulfoxide (DMSO), toluene, n-hexane, dichloromethane；
(2) extract solution obtained by step (1) is carried out acidic silica gel post, acidic alumina silica gel column purification by (2) successively, obtains net
Change liquid；
(3) scavenging solution obtained by step (2) is blown into concentration through nitrogen, is added13C-PCDD/Fs internal standards, obtain Instrumental Analysis purification
Sample.
2. milk sample pre-treating method as claimed in claim 1, it is characterised in that in the step (1), the milk sample
Product are commercially available liquid milk or natural liquid milk.
3. milk sample pre-treating method as claimed in claim 1, it is characterised in that the described step for hydrolyzing milk sample
Suddenly, specifically include：
Milk sample is mixed with alkaline protease solution, 3~5h is hydrolyzed under conditions of 40~55 DEG C, pH=8.5~10.0；
Add alkaline solution and hydrolyze 15~35min at 70~80 DEG C, add Organic Alcohol concussion and be well mixed, wherein, the alkalescence
The mass fraction of protein enzyme solution neutral and alkali protease is 1~5%；The body of the alkaline protease solution and the milk sample
Product is than being 1:10~20；The alkaline solution includes the one kind in ammonia spirit, sodium hydrate aqueous solution, potassium hydroxide aqueous solution
Or it is several；The Organic Alcohol includes one or more in ethanol, methyl alcohol, butanediol, propyl alcohol.
4. milk sample pre-treating method as claimed in claim 1, it is characterised in that in the step (1), described addition
The step of extraction of first organic solvent, separation water outlet phase and the first organic phase, specifically include：
The first organic solvent, 3~10min of oscillation extraction is added to separate water outlet phase and the first organic phase.
5. milk sample pre-treating method as claimed in claim 1, it is characterised in that in the step (1), to the first water phase
Middle addition the second organic solvent extraction, the step of isolate the second water phase and Second Organic Phase, specifically includes：
The second water phase and Second Organic Phase are isolated to the second organic solvent, 3~10min of oscillation extraction is added in the first water phase.
6. milk sample pre-treating method as claimed in claim 1, it is characterised in that in the step (1), it is described by institute
State concentrate to be well mixed with mix reagent, centrifugation, removal the concentrated sulfuric acid layer, be obtained extract solution the step of, specifically include：
By the concentrate and organic solvent, inorganic acid solution vortex shaken well, then centrifuge is used with 2000~4000r/
The centrifugation rate of min is centrifuged 0.5~4min, and the concentrated sulfuric acid of lower floor is then sucked with dropper, and extract solution is obtained, wherein, it is described to have
Machine solvent includes one or more in ether, acetone, dimethyl sulfoxide (DMSO), toluene, n-hexane, dichloromethane；The inorganic acid
Solution is concentrated sulfuric acid solution；The organic solvent is 1~10 with the volume ratio of the concentrate:1；The inorganic acid solution and institute
The volume ratio for stating concentrate is 1~10:1.
7. milk sample pre-treating method as claimed in claim 1, it is characterised in that the step (2), specifically includes：
The extract solution obtained by step (1) is purified using the acidic silica gel post after being washed with the first pre- eluent pre-leaching, is passed through
After first eluent drip washing, the first scavenging solution is concentrated to give；The scavenging solution is transferred to by pre- drip washing using the second eluent
In the acidic alumina silicagel column for the treatment of, after carrying out drip washing using the second eluent, by the anti-mistake of acidic alumina silicagel column
After coming, using the 3rd eluent drip washing, and collection obtains the second scavenging solution；
Wherein, the described first pre- eluent includes one or more in ether, toluene, n-hexane and dichloromethane；Described
One eluent includes one or more in ether, toluene, n-hexane and dichloromethane；Second eluent include just oneself
One or more in alkane, dichloromethane, methyl alcohol, toluene；3rd eluent is included in n-hexane, dichloromethane, toluene
One or more.
8. milk sample pre-treating method as claimed in claim 7, it is characterised in that in the step (2), the pre-leaching
The acidic alumina silicagel column for washing treatment is to be obtained using following methods：
Pre- drip washing for the first time is carried out to the acidic alumina silicagel column using the second pre- eluent, then using the 3rd pre- eluent
Second pre- drip washing is carried out to the acidic alumina silicagel column, then using the 4th pre- eluent to the acidic oxidation aluminum silica gel
Post carries out the pre- drip washing of third time, then the 4th pre- drip washing is carried out to the acidic alumina silicagel column using the 5th pre- eluent,
Wherein, the described second pre- eluent, the 3rd pre- eluent, the 4th pre- eluent, the 5th pre- eluent are identical or different pre-
Drip washing organic solvent, wherein, the pre- drip washing organic solvent includes the one kind or several in n-hexane, dichloromethane, methyl alcohol, toluene
Kind.
9. milk sample pre-treating method as claimed in claim 1, it is characterised in that the step (3), specifically includes：
Scavenging solution obtained by step (2) is transferred in sample bottle, nitrogen is blown to do, is transferred in the sample bottle that capacity is 1.5mL,
10~14h is placed, is added13C-PCDD/Fs internal standard constant volumes, the purification sample of prepared Instrumental Analysis.
10. a kind of assay method of milk sample Zhong bioxin, it is characterised in that comprise the following steps：
Take and purify sample obtained in the milk sample pre-treating method as described in claim 1~9 is any, using isotopic dilution
High resolution spectral estimation combined instrument carries out Instrumental Analysis measure to the purification sample, in drawing milk sample
The measurement result of content of dioxin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710152294.3A CN106908303A (en) | 2017-03-15 | 2017-03-15 | A kind of pre-treating method of milk sample is Ji the assay method of bioxin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710152294.3A CN106908303A (en) | 2017-03-15 | 2017-03-15 | A kind of pre-treating method of milk sample is Ji the assay method of bioxin |
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