CN106501431A - A kind of silica gel chromatographic column and its detection method for detecting Polychlorinated biphenyls in gas chromatograph-mass spectrometer - Google Patents
A kind of silica gel chromatographic column and its detection method for detecting Polychlorinated biphenyls in gas chromatograph-mass spectrometer Download PDFInfo
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- CN106501431A CN106501431A CN201610915107.8A CN201610915107A CN106501431A CN 106501431 A CN106501431 A CN 106501431A CN 201610915107 A CN201610915107 A CN 201610915107A CN 106501431 A CN106501431 A CN 106501431A
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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
- G01N30/60—Construction of the column
- G01N30/6034—Construction of the column joining multiple columns
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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
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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
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- G01N30/08—Preparation using an enricher
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Abstract
The invention provides a kind of silica gel chromatographic column for detecting Polychlorinated biphenyls in gas chromatograph-mass spectrometer, the silica gel chromatographic column purifies chromatographic column, compound silica gel purification chromatographic column and alkali alumina purification chromatographic column by acidic silica gel and constitutes, and specifically discloses the preparation method of each chromatographic column.The present invention additionally provides a kind of method using Polychlorinated biphenyls in above-mentioned silica gel chromatography post detection milk product simultaneously, the modified silica-gel superior performance prepared in the method, and in the method implant fill order with strong points, fill order is reasonable, therefore, multilamellar silica gel purification chromatographic column prepared by the method can overcome the matrix effect of complexity, the plurality of impurities in the organic pollution matrix of class containing polychlorinated biphenyl can be removed, eliminate the interference produced by follow-up polychlorinated biphenyl class organic contamination analyte detection, so as to improve gas chromatograph-mass spectrometer accurate quantitative analysis polychlorinated biphenyl, simple to operate, reproducible, high precision.
Description
Technical field
The present invention relates to a kind of preparation method of the silica gel chromatographic column of gas chromatograph-mass spectrometer detection Polychlorinated biphenyls, with
And the detection method of the silica gel chromatography post detection Polychlorinated biphenyls, detection technique field in milk product.
Background technology
Chlordiphenyl (PCBs) be a class with biphenyl be raw material under metallic catalyst effect, the chloro virtue of high-temp chlorination synthesis
Hydrocarbon.According to the difference that chlorine atom replaces number and the position of substitution, PCBs has 209 kinds of homologues.Due to PCBs have good
The advantages of chemical inertness, heat resistance, incombustible, low-steam pressure and high-k, therefore it is used for heat exchanging agent, lubrication
Dielectric, plasticizer in agent, transformator and capacitor, paraffin expand agent, adhesive, organic diluent, dedusting agent, parasite killing
The important chemical products such as agent, cutting oil, pressure-sensitive carbon paper and fire retardant, and it is widely used in power industry, plastic processing
The fields such as industry, chemical industry and printing.PCBs has persistency and extended residual, once it is discharged in environment, it is difficult to be degraded, because
Can there is many decades or longer time in this in the media such as water body, soil, in PCBs structures, contained chlorine atom has which
The characteristics of having low aqueous solubility and fat-solubility, it is thus possible to which biological accumulation occurs in fatty tissue, and by the life of food chain
Thing amplification reaches the concentration of poisoning.
The organism kinds of milk product are various, and the matrix residing for this kind of organic pollution is also sufficiently complex, if impurity
Without process, then larger interference can be produced to the testing result of follow-up organic pollution, affect the result of detection.This method is adopted
Silica gel chromatography column purification, according to material, absorption affinity on silica gel is different and separated, generally the larger thing of polarity
Easily by silica gel adsorption, the weaker material of polarity is difficult by silica gel adsorption matter, whole chromatography process is Adsorption and desorption, adsorb again,
Desorption process, adopts acidic silica gel column purification, compound silica gel column purification, alkali alumina column purification successively again.Due to ordinary silicon
The interference produced by the detection of follow-up polychlorinated biphenyl of glue chromatographic column is very big.Therefore, prepared by common silica gel chromatographic column
Silica gel chromatographic column prepared by method cannot be used for polychlorinated biphenyl compound class organic pollution in detection of complex matrix.
Content of the invention
This method purpose is to solve the content analysis method for improving polychlorinated biphenyl, reduce impurity in sample
And the interference of exogenous impurity, so as to improve gas chromatograph-mass spectrometer accurate quantitative analysis polychlorinated biphenyl, there is provided
The assay method of the content of polychlorinated biphenyl in a kind of detection milk product of simple to operate, reproducible, high precision.
Realize technical scheme that above-mentioned purpose of the present invention adopted for：
A kind of silica gel chromatographic column for detecting Polychlorinated biphenyls in gas chromatograph-mass spectrometer, the silica gel chromatographic column is by acid
Property silica gel purification chromatographic column, compound silica gel purify chromatographic column and alkali alumina and purify chromatographic column composition, the preparation of each chromatographic column
Method is as follows：
(1), each implant component is weighed according to following mass parts：
A acidic silica gels purify chromatographic column：1. 2～4 parts of activated silica gel；2. 4～6 parts of acidic silica gel；3. activated silica gel 1
～2 parts；4. 2～4 parts of anhydrous sodium sulfate；
B compound silica gels purify chromatographic column：1. 1～3 part of silver nitrate silica gel；2. 1～2 part of activated silica gel；3. alkaline silica gel
2～4 parts；4. 1～2 part of activated silica gel；5. 3～6 parts of acidic silica gel；6. 2～4 parts of activated silica gel；7. anhydrous sodium sulfate 2～
4 parts；
C alkali aluminas purify chromatographic column：1. 1～3 part of alkali alumina；2. 2～4 parts of anhydrous sodium sulfate；
(2), the chromatographic column of three dried and cleans is taken, and chromatograph column internal diameter is 1 with length ratio:8～10, according to step (1)
Each implant is poured into by the order of middle each component successively successively from the upper port of chromatographic column, and tri- kinds of proportionings of A, B and C correspond to one respectively
Individual chromatographic column, after every kind of implant is poured into from the upper port of chromatographic column, allows the implant for pouring into tile uniform, and chromatograph column wall
Non-filler adheres to, and after the completion of all implant fills, opens the rotary-piston of chromatographic column lower end, connects in the lower port of chromatographic column
Enter vacuum pump, then open vacuum pump, beat the outer wall of chromatographic column in evacuation, when chromatographic column inner stuffing height no longer
During decline, continue evacuation after at least 3 minutes, screw chromatographic column lower end piston；
(3) dichloromethane is slowly injected chromatographic column from chromatographic column upper port along chromatograph column wall, chromatographic column lower end is opened
Piston, make the flow velocity of the liquid of inflow chromatographic column maximum, make all implants all wetted and bubble-free, when in chromatographic column
Each implant clear layer, form for the moment, stop injection dichloromethane, finally with enough normal hexane elution chromatography posts, wash
Take off the piston that chromatographic column lower end is closed after finishing.
The preparation method of the activated silica gel is as follows：Filling is adopted normal hexane and eluent methylene chloride twice with silica gel,
The solvent volume for using every time is the twice of silica gel volume, after drip washing, silica gel is transferred in flask, covers bottleneck with aluminium foil and put
In baking oven 50 DEG C be baked to dry, then heat at 150～250 DEG C toast 8h～12h, after cooling load ground reagent bottle
In, preserve in exsiccator, that is, activated silica gel is obtained.
The preparation method of the silver nitrate silica gel is as follows：4～7g silver nitrate is dissolved in 15～25mL deionized waters, by
It is added dropwise in 50g activated silica gels, shakes to after without block, loads in brown ground reagent bottle, preserve in exsiccator, that is, be obtained
Silver nitrate silica gel.
The acidic silica gel is sulphuric acid silica gel, and the preparation method of sulphuric acid silica gel is as follows：Press sulphuric acid：Silica gel=8:8～12
Mass ratio, is added drop-wise to the concentrated sulphuric acid that mass percent concentration is 95%～97% in silica gel under agitation, per Deca one
Drop concentrated sulphuric acid will stir to after being uniformly dispersed again the next drop concentrated sulphuric acid of Deca, after concentrated sulphuric acid completion of dropping, will be dense with agitator
Sulphuric acid is finished to reaction with the mixed system sustained oscillation of silica gel, is loaded in ground reagent bottle, is preserved in exsiccator, that is, sulfur is obtained
Sour silica gel.
The alkaline silica gel is sodium hydroxide silica gel, and the preparation method of sodium hydroxide silica gel is as follows：Press sodium hydroxide solution：
Silica gel=1:2～3 mass ratio, under agitation by concentration for 0.8mol/L～1.2mol/L sodium hydroxide solution Deca
To in silica gel, per Deca one, drop sodium hydroxide solution will stir to after being uniformly dispersed again the next drop sodium hydroxide solution of Deca,
After completion of dropping, the mixed system sustained oscillation with agitator by sodium hydroxide solution with silica gel is finished to reaction, loads ground
In reagent bottle, preserve in exsiccator, that is, sodium hydroxide silica gel is obtained.
The time that is shaken with agitator in above-mentioned steps is 6～8 hours.
The alkali alumina is using the pretreated alkali alumina of following methods：By alkali alumina 600～
At 700 DEG C after 4～8h of baking, load in ground reagent bottle, preserve in exsiccator.
The anhydrous sodium sulfate is anhydrous sodium sulfate pretreated through following methods：By anhydrous sodium sulfate using just oneself
Alkane and eluent methylene chloride 2～5 times, twice of the solvent volume for using every time for anhydrous sodium sulfate volume, after drip washing, will be anhydrous
Sodium sulfate is transferred in flask, be baked at 50 DEG C dry, then at 150～250 DEG C toast 8h～12h, after cooling dry
Preserve in device.
The present invention additionally provides a kind of method using Polychlorinated biphenyls in above-mentioned silica gel chromatography post detection milk product simultaneously, wraps
Include following steps：
(1), milk product sample is put in refrigerator and freezes, to be frozen completely after, frozen dairy product is put into freezer dryer,
- 20 DEG C～-40 DEG C of freezer dryer precooling temperature, -30 DEG C～-40 DEG C of lyophilization temperature, by lyophilizing after sample load
Surname extraction filter cylinder simultaneously places soxhlet type apparatus, and 5h～8h is extracted in water-bath, and siphonage time is 3～4 times/h, and the sample of concentration is obtained
Product extracting solution；
(2), rinse acid silica gel purification chromatographic column, compound silica gel respectively with the mixed liquor of normal hexane and dichloromethane to purify
Chromatographic column and alkali alumina purify chromatographic column, and the flow velocity of flushing is maximum, makes liquid level equal with silica gel；
(3) sample extracting solution of concentration is injected into acidic silica gel, to purify in chromatographic column, normal hexane eluting is used, control is solidifying
The flow velocity of glue post is 1.0～3.0mL/min, and eluent is concentrated into 0.5～1.5mL；
To purify through acidic silica gel the extracting solution after chromatograph column purification be injected into compound silica gel purify chromatographic column in, with just oneself
Alkane and dichloromethane are according to 97:1～97:The pre- drip washing of the mixed solution of 4 volume ratio；Chromatograph will be purified again through acidic silica gel
After column purification, concentrate eluant is fully transferred in compound silica gel purification chromatographic column；Add when liquid level is down to anhydrous sodium sulfate layer
Normal hexane and dichloromethane are according to 97:1～97:The mixed solution of 4 volume ratio carries out eluting, and eluent is concentrated into 0.5
～1.0mL；
The extracting solution for purifying chromatograph column purification through compound silica gel is injected into alkali alumina to purify in chromatographic column, will be through multiple
After closing silica gel purification chromatograph column purification, concentrate eluant is fully transferred in alkali alumina purification chromatographic column, when liquid level is down to nothing
Add normal hexane eluting alkali alumina to purify chromatographic column during aqueous sodium persulfate layer, two are added when liquid level is down to anhydrous sodium sulfate layer
Chloromethanes and the mixed solution eluting of normal hexane, then eluent is concentrated near dry；
(4), the concentrate eluant in step (3) is analyzed using gas chromatograph-mass spectrometer；
(5), quantitative analyses, using double internal standard quanitations, 8 kinds of Polychlorinated biphenyls Isotopic Internal Standards (13C12-labelled
PCBs 28,52,118,153,180,202,206 and 209) He 2 kind of Polychlorinated biphenyls isotope reclaim internal standard (13C12-PCB101、13C12- PCB194) quantitative, Isotopic Internal Standard is used for calibrating the determinand of 33 kinds of Polychlorinated biphenyls.
In above-mentioned steps (4), the operating parameter of gas chromatograph-mass spectrometer is：Sampling (5%- phenyl)-methyl polysilicon oxygen
The capillary chromatographic column of alkane filler, the dottle pin of high temperature resistant low-bleed and graphite grazing pad, deactivation are filled with glass cotton shunting in processing
With regardless of flowing universal bushing pipe；The program of column oven is：Maintain 2 minutes at 100 DEG C first, then the heating rate with 15 DEG C/min
180 DEG C are raised to from 100 DEG C；Then 240 DEG C are raised to the heating rate of 3 DEG C/min from 180 DEG C；Finally with the intensification of 10 DEG C/min
Speed is raised to 285 DEG C from 240 DEG C, and maintains 10min at 285 DEG C；Total run time is 42min；Carrier gas uses purity>
99.999% high-purity helium, constant flow rate are 1mL/min；The sample size of sample and standard liquid is 2.0 μ L, using Splitless injecting samples
Pattern, injector temperature are 300 DEG C；Mass spectrometer parameters are：Ionization pattern, electron bombardment ionization source (EI), energy is 70eV；Ion source
Temperature is 230 DEG C, and 150 DEG C of level Four bar temperature, transmission line temperature are 300 DEG C, and solvent delay is 4min.
Compared with prior art, its advantage and advantage are the present invention：
In the method prepare modified silica-gel superior performance, and in the method implant fill order with strong points,
Rationally, therefore, multilamellar silica gel purification chromatographic column prepared by the method can overcome the matrix effect of complexity to fill order, can remove and contain
Plurality of impurities in polychlorinated biphenyl class organic pollution matrix, eliminating has to follow-up polychlorinated biphenyl class
The interference that organic pollutants detection is produced.Importantly, multilamellar silica gel purification chromatographic column prepared by the method is simple to operate, through examination
Test and show, the organic pollution such as multilamellar silica gel chromatography post detection polychlorinated biphenyl prepared using the method has sensitive
The advantage that degree is high, accuracy is high, favorable reproducibility and the response rate are high.
Specific embodiment
Silica gel chromatographic column for detection Polychlorinated biphenyls in gas chromatograph-mass spectrometer provided by the present invention is by acidity
Silica gel purification chromatographic column, compound silica gel purify chromatographic column and alkali alumina purifies chromatographic column composition, the following institute of its preparation method
Show：
(1), each implant component is weighed according to following mass parts：
A acidic silica gels purify chromatographic column：1. 2～4 parts of activated silica gel；2. 4～6 parts of acidic silica gel；3. activated silica gel 1
～2 parts；4. 2～4 parts of anhydrous sodium sulfate；
B compound silica gels purify chromatographic column：1. 1～3 part of silver nitrate silica gel；2. 1～2 part of activated silica gel；3. alkaline silica gel
2～4 parts；4. 1～2 part of activated silica gel；5. 3～6 parts of acidic silica gel；6. 2～4 parts of activated silica gel；7. anhydrous sodium sulfate 2～
4 parts；
C alkali aluminas purify chromatographic column：1. 1～3 part of alkali alumina；2. 2～4 parts of anhydrous sodium sulfate；
Wherein：The preparation method of the silver nitrate silica gel is as follows：4～7g silver nitrate is dissolved in 15～25mL deionized waters
In, be added dropwise in 50g activated silica gels, 6～8 hours are vibrated to after without block with agitator, load brown ground reagent bottle
In, preserve in exsiccator, that is, silver nitrate silica gel is obtained.
The acidic silica gel is sulphuric acid silica gel, and the preparation method of sulphuric acid silica gel is as follows：Press sulphuric acid：Silica gel=8:8～12
Mass ratio, is added drop-wise to the concentrated sulphuric acid that mass percent concentration is 95%～97% in silica gel under agitation, per Deca one
Drop concentrated sulphuric acid will stir to after being uniformly dispersed again the next drop concentrated sulphuric acid of Deca, after concentrated sulphuric acid completion of dropping, will be dense with agitator
Sulphuric acid is finished with the mixed system sustained oscillation of silica gel for 6～8 hours to reaction, is loaded in ground reagent bottle, is preserved in exsiccator,
Sulphuric acid silica gel is obtained.
The alkaline silica gel is sodium hydroxide silica gel, and the preparation method of sodium hydroxide silica gel is as follows：Press sodium hydroxide solution：
Silica gel=1:2～3 mass ratio, under agitation by concentration for 0.8mol/L～1.2mol/L sodium hydroxide solution Deca
To in silica gel, per Deca one, drop sodium hydroxide solution will stir to after being uniformly dispersed again the next drop sodium hydroxide solution of Deca,
After completion of dropping, the mixed system sustained oscillation with agitator by sodium hydroxide solution with silica gel is finished for 6～8 hours to reaction,
Load in ground reagent bottle, preserve in exsiccator, that is, sodium hydroxide silica gel is obtained.
The preparation method of the activated silica gel is as follows：Filling is adopted normal hexane and eluent methylene chloride twice with silica gel,
The solvent volume for using every time is the twice of silica gel volume, after drip washing, silica gel is transferred in flask, covers bottleneck with aluminium foil and put
In baking oven 50 DEG C be baked to dry, then heat at 150～250 DEG C toast 8h～12h, after cooling load ground reagent bottle
In, preserve in exsiccator, that is, activated silica gel is obtained.
The alkali alumina is using the pretreated alkali alumina of following methods：By alkali alumina 600～
At 700 DEG C after 4～8h of baking, load in ground reagent bottle, preserve in exsiccator.
The anhydrous sodium sulfate is anhydrous sodium sulfate pretreated through following methods：By anhydrous sodium sulfate using just oneself
Alkane and eluent methylene chloride 2～5 times, twice of the solvent volume for using every time for anhydrous sodium sulfate volume, after drip washing, will be anhydrous
Sodium sulfate is transferred in flask, be baked at 50 DEG C dry, then at 150～250 DEG C toast 8h～12h, after cooling dry
Preserve in device.
(2), the chromatographic column of three dried and cleans is taken, and chromatograph column internal diameter is 1 with length ratio:8～10, according to step (1)
Each implant is poured into by the order of middle each component successively successively from the upper port of chromatographic column, and tri- kinds of proportionings of A, B and C correspond to one respectively
Individual chromatographic column, after every kind of implant is poured into from the upper port of chromatographic column, allows the implant for pouring into tile uniform, and chromatograph column wall
Non-filler adheres to, and after the completion of all implant fills, opens the rotary-piston of chromatographic column lower end, connects in the lower port of chromatographic column
Enter vacuum pump, then open vacuum pump, beat the outer wall of chromatographic column in evacuation, when chromatographic column inner stuffing height no longer
During decline, continue evacuation after at least 3 minutes, screw chromatographic column lower end piston；
(3) dichloromethane is slowly injected chromatographic column from chromatographic column upper port along chromatograph column wall, chromatographic column lower end is opened
Piston, make the flow velocity of the liquid of inflow chromatographic column maximum, make all implants all wetted and bubble-free, when in chromatographic column
Each implant clear layer, form for the moment, stop injection dichloromethane, finally with enough normal hexane elution chromatography posts, wash
Take off the piston that chromatographic column lower end is closed after finishing.
The feasibility of the silica gel chromatographic column of the detection Polychlorinated biphenyls for providing for the inspection present invention below and superiority, in breast system
Product add polychlorinated biphenyl compound standard solution, the sample extraction that will be provided using the present invention in the milk product containing standard solution
Method and silica gel purification chromatographic column are separated, and detect the content of the polychlorinated biphenyl compound in milk product, according to the knot of detection
Fruit concentration and the difference of known organic concentration, enter to sample extraction, purification and eluting feasibility and superiority in the method
Row assessment.
Embodiment
In the present embodiment, acidic silica gel purification chromatographic column filling is as follows：Glass column bottom with glass cotton closure after from bottom to
4g activated silica gel, 10g acidifying silica gel, 2g activated silica gel, 4g anhydrous sodium sulfate are inserted successively in top.
It is as follows that compound silica gel purifies chromatographic column filling：According to inserting from bottom to top after the closure of glass column bottom glass cotton
1.5g silver nitrate silica gel, 1g activated silica gels, 2g alkaline silica gels, 1g activated silica gels, 4g acidifying silica gel, 2g activated silica gels, 2g are anhydrous
Sodium sulfate.
It is as follows that alkali alumina purifies chromatographic column filling：According to filling out from bottom to top after the closure of glass column bottom glass cotton
Enter alkali alumina, 2g anhydrous sodium sulfate of the 2.5g through overbaking.
Take 4.0mL mixed standard solution (PCB 1, PCB 3, PCB 4, PCB 15, PCB 19, PCB 23, PCB 34,
PCB 37、PCB 54、PCB 77、PCB 81、PCB 104、PCB 105、PCB 114、PCB 118、PCB 123、PCB 126、
PCB 155、PCB 156、PCB 157、PCB 167、PCB 169、PCB 170、PCB 180、PCB 182、PCB 187、PCB
188th, PCB 189, PCB 202, PCB 205, PCB 206, PCB 208 and PCB 209, concentration are 20 μ g mL-1, Polychlorinated biphenyls
Isotopic Internal Standard, comprising homologue13C12- labelled PCBs 28,52,118,153,180,202,206 and 209, concentration is
2μg mL-1) in the milk product sample that is added separately to, the milk product sample had carried out following pre-treatment in advance：Milk product
Sample is put in refrigerator (- 18 DEG C) freezings, to be frozen completely after, frozen dairy product is put into freezer dryer, freezer dryer
- 20 DEG C～-40 DEG C of precooling temperature, -30 DEG C～-40 DEG C of lyophilization temperature.Then surname extraction is carried out, by pretreating specimen
5.0g loads surname extraction filter cylinder and places soxhlet type apparatus, extracts 5h-8h in 60 DEG C of water-baths, and siphonage time is 3-4 time/h,
The sample extracting solution of concentration is obtained.
Acid silica gel purification chromatographic column, compound silica gel are rinsed with normal hexane respectively with the mixed liquor of dichloromethane and purify chromatograph
Post and alkali alumina purify chromatographic column, and the flow velocity of flushing is maximum, makes liquid level equal with silica gel.
Sample 0.5～1.5mL extracting solution of concentration is injected into acidic silica gel to purify in chromatographic column, with 80～120mL just
Hex, the flow velocity for controlling gel column is 1.0～3.0mL/min, and eluent is concentrated into 0.5～1.5mL；
The extracting solution after chromatograph column purification will be purified through acidic silica gel be injected into compound silica gel purify in chromatographic column, with 20～
50mL normal hexane and dichloromethane are according to 97:1～97:The pre- drip washing of the mixed solution of 4 volume ratio；Again will be through acidic silica gel
After purifying chromatograph column purification, concentrate eluant is fully transferred in compound silica gel purification chromatographic column, rinses eggplant with 3～6mL normal hexane
Shape bottle 3 times～4 times, washing liquid is transferred on post.40～80mL normal hexane and dichloro is added when liquid level is down to anhydrous sodium sulfate layer
Methane is according to 97:1～97:The mixed solution of 4 volume ratio carries out eluting, and eluent is concentrated into 0.5～1.0mL.
The extracting solution for purifying chromatograph column purification through compound silica gel is injected into alkali alumina to purify in chromatographic column, will be through multiple
Close silica gel purification chromatograph column purification after concentrate eluant be fully transferred to alkali alumina purify chromatographic column on, with 3～6mL just oneself
Alkane flushing eggplant-shape bottle 3 times～4 times, washing liquid is transferred on post.When liquid level is down to anhydrous sodium sulfate layer add 20～50mL just oneself
Alkane eluting alkali alumina purifies chromatographic column, adds the mixing of dichloromethane and normal hexane when liquid level is down to anhydrous sodium sulfate layer
Eluant solution, then eluent is concentrated near dry.
Above-mentioned concentrate eluant is analyzed using gas chromatograph-mass spectrometer, (5%- phenyl)-methyl polysilicon oxygen of sampling
The capillary chromatographic column of alkane filler, the dottle pin of high temperature resistant low-bleed and graphite grazing pad, deactivation are filled with a small amount of glass cotton in processing
Shunt and regardless of the universal bushing pipe of stream；The program of column oven is：Maintain 2 minutes at 100 DEG C first, then with 15 DEG C of min-1Intensification
Speed is raised to 180 DEG C from 100 DEG C；Then with 3 DEG C of min-1Heating rate be raised to 240 DEG C from 180 DEG C；Finally with 10 DEG C of min-1
Heating rate be raised to 285 DEG C from 240 DEG C, and 285 DEG C maintain 10min.Total run time is 42min.Carrier gas is using high
Pure helium (purity>99.999%), constant flow rate is 1mL min-1.The sample size of sample and standard liquid is 2.0 μ L, using not shunting
Sample introduction pattern, injector temperature are 300 DEG C.Make all of polychlorinated biphenyl in matrix completely separable.Mass spectrometry parameters are：
Ionization pattern, electron bombardment ionization source (EI), energy is 70eV.Ion source temperature is 230 DEG C, 150 DEG C of level Four bar temperature, transmission line temperature
Spend for 300 DEG C, solvent delay is 4min.
Quantitative analyses are finally carried out, using double internal standard quanitations, 8 kinds of Polychlorinated biphenyls Isotopic Internal Standards (13C12-labelled
PCBs 28,52,118,153,180,202,206 and 209) He 2 kind of Polychlorinated biphenyls isotope reclaim internal standard (13C12-PCB101、13C12- PCB194) quantitative, Isotopic Internal Standard is used for calibrating the determinand of 33 kinds of Polychlorinated biphenyls.After being computed in the present embodiment
Testing result is as follows：In milk product, the response rate of the polychlorinated diphenyl ether (33 kinds) under the mark-on level of 20 μ g/kg is：88%～
105%.
The result of the above-mentioned response rate is met in european union directive (European Union document 2002/657/EC)
Related request.During 10 μ g/kg≤100 μ g/kg of P ＜, the response rate need to be in the range of 80%～110%, and P refers to the concentration of determinand.
Result according to example detection can be seen that the response rate of each persistent organism and all meet european union directive
Related request in (European Union document 2002/657/EC), illustrates the multilamellar silicon provided using the present invention
Glue purifies the preparation of chromatographic column and the multilamellar silica gel chromatographic column of fill method preparation can be used for detecting in environmental sample there there is persistency
The content of machine thing, and sensitivity is high, accuracy is high, favorable reproducibility and the response rate are high.
Claims (10)
1. a kind of in gas chromatograph-mass spectrometer detect Polychlorinated biphenyls silica gel chromatographic column, it is characterised in that the silica gel color
Spectrum post purifies chromatographic column, compound silica gel purification chromatographic column and alkali alumina purification chromatographic column by acidic silica gel and constitutes, each chromatograph
The preparation method of post is as follows：
(1), each implant component is weighed according to following mass parts：
A acidic silica gels purify chromatographic column：1. 2～4 parts of activated silica gel；2. 4～6 parts of acidic silica gel；3. 1～2 part of activated silica gel；④
2～4 parts of anhydrous sodium sulfate；
B compound silica gels purify chromatographic column：1. 1～3 part of silver nitrate silica gel；2. 1～2 part of activated silica gel；3. 2～4 parts of alkaline silica gel；
4. 1～2 part of activated silica gel；5. 3～6 parts of acidic silica gel；6. 2～4 parts of activated silica gel；7. 2～4 parts of anhydrous sodium sulfate；
C alkali aluminas purify chromatographic column：1. 1～3 part of alkali alumina；2. 2～4 parts of anhydrous sodium sulfate；
(2), the chromatographic column of three dried and cleans is taken, and chromatograph column internal diameter is 1 with length ratio:8～10, according to each in step (1)
Each implant is poured into by the order of component successively successively from the upper port of chromatographic column, and tri- kinds of proportionings of A, B and C correspond to a color respectively
Spectrum post, after every kind of implant is poured into from the upper port of chromatographic column, allows the implant that pours into tile uniform, and chromatograph column wall is without filling out
Thing attachment is filled, the rotary-piston of chromatographic column lower end after the completion of all implant fills, is opened, is accessed in the lower port of chromatographic column true
Empty pump, then opens vacuum pump, beats the outer wall of chromatographic column in evacuation, when the height of chromatographic column inner stuffing no longer declines
When, continue evacuation after at least 3 minutes, screw chromatographic column lower end piston；
(3) dichloromethane is slowly injected chromatographic column from chromatographic column upper port along chromatograph column wall, the work of chromatographic column lower end is opened
Plug, makes the flow velocity of the liquid of inflow chromatographic column maximum, makes all implants all wetted and bubble-free, when respectively filling out in chromatographic column
Fill thing clear layer, form for the moment, stop injection dichloromethane, finally with enough normal hexane elution chromatography posts, eluting is complete
The piston of chromatographic column lower end is closed after finishing.
2. according to claim 1 in gas chromatograph-mass spectrometer detect Polychlorinated biphenyls silica gel chromatographic column, its
It is characterised by：The preparation method of the activated silica gel is as follows：Filling is adopted normal hexane and eluent methylene chloride twice with silica gel,
The solvent volume for using every time is the twice of silica gel volume, after drip washing, silica gel is transferred in flask, covers bottleneck with aluminium foil and put
In baking oven 50 DEG C be baked to dry, then heat at 150～250 DEG C toast 8h～12h, after cooling load ground reagent bottle
In, preserve in exsiccator, that is, activated silica gel is obtained.
3. according to claim 1 in gas chromatograph-mass spectrometer detect Polychlorinated biphenyls silica gel chromatographic column, its
It is characterised by：The preparation method of the silver nitrate silica gel is as follows：4～7g silver nitrate is dissolved in 15～25mL deionized waters,
It is added dropwise in 50g activated silica gels, shakes to after without block, loads in brown ground reagent bottle, preserve in exsiccator, that is, make
Obtain silver nitrate silica gel.
4. according to claim 1 in gas chromatograph-mass spectrometer detect Polychlorinated biphenyls silica gel chromatographic column, its
It is characterised by：The acidic silica gel is sulphuric acid silica gel, and the preparation method of sulphuric acid silica gel is as follows：Press sulphuric acid：Silica gel=8:8～12
Mass ratio, is added drop-wise to the concentrated sulphuric acid that mass percent concentration is 95%～97% in silica gel under agitation, per Deca one
Drop concentrated sulphuric acid will stir to after being uniformly dispersed again the next drop concentrated sulphuric acid of Deca, after concentrated sulphuric acid completion of dropping, will be dense with agitator
Sulphuric acid is finished to reaction with the mixed system sustained oscillation of silica gel, is loaded in ground reagent bottle, is preserved in exsiccator, that is, sulfur is obtained
Sour silica gel.
5. according to claim 1 in gas chromatograph-mass spectrometer detect Polychlorinated biphenyls silica gel chromatographic column, its
It is characterised by：The alkaline silica gel is sodium hydroxide silica gel, and the preparation method of sodium hydroxide silica gel is as follows：Press sodium hydroxide molten
Liquid：Silica gel=1:2～3 mass ratio, under agitation by concentration for 0.8mol/L～1.2mol/L sodium hydroxide solution
It is added drop-wise in silica gel, it is molten that the every drop of Deca one sodium hydroxide solution will stir to after being uniformly dispersed again the next drop sodium hydroxide of Deca
Liquid, after completion of dropping, the mixed system sustained oscillation with agitator by sodium hydroxide solution with silica gel is finished to reaction, loads mill
In opening reagent bottle, preserve in exsiccator, that is, sodium hydroxide silica gel is obtained.
6. according to claim 3 or 4 or 5 for gas chromatograph-mass spectrometer in detect Polychlorinated biphenyls silica gel chromatography
Post, it is characterised in that：It is 6～8 hours with the time that agitator shakes.
7. according to claim 1 in gas chromatograph-mass spectrometer detect Polychlorinated biphenyls silica gel chromatographic column, its
It is characterised by：The alkali alumina is using the pretreated alkali alumina of following methods：By alkali alumina 600～
At 700 DEG C after 4～8h of baking, load in ground reagent bottle, preserve in exsiccator.
8. according to claim 1 in gas chromatograph-mass spectrometer detect Polychlorinated biphenyls silica gel chromatographic column, its
It is characterised by：The anhydrous sodium sulfate is anhydrous sodium sulfate pretreated through following methods：By anhydrous sodium sulfate using just
Hexane and eluent methylene chloride 2～5 times, twice of the solvent volume for using every time for anhydrous sodium sulfate volume, after drip washing, by nothing
Aqueous sodium persulfate is transferred in flask, be baked at 50 DEG C dry, then at 150～250 DEG C toast 8h～12h, after cooling do
Preserve in dry device.
9. in a kind of employing claim 1-8 in arbitrary described silica gel chromatography post detection milk product Polychlorinated biphenyls method, its
It is characterised by comprising the following steps：
(1), milk product sample is put in refrigerator and freezes, to be frozen completely after, frozen dairy product is put into freezer dryer, is freezed
- 20 DEG C～-40 DEG C of drying machine precooling temperature, -30 DEG C～-40 DEG C of lyophilization temperature, by lyophilizing after sample load Soxhlet
Extract filter cylinder and place soxhlet type apparatus, 5h～8h is extracted in water-bath, siphonage time is 3～4 times/h, the sample for concentrating is obtained and carries
Take liquid；
(2) acid silica gel purification chromatographic column, compound silica gel are rinsed respectively with the mixed liquor of normal hexane and dichloromethane, and purifies chromatograph
Post and alkali alumina purify chromatographic column, and the flow velocity of flushing is maximum, makes liquid level equal with silica gel；
(3) sample extracting solution of concentration is injected into acidic silica gel, to purify in chromatographic column, normal hexane eluting is used, gel column is controlled
Flow velocity be 1.0～3.0mL/min, eluent is concentrated into 0.5～1.5mL；
The extracting solution after chromatograph column purification will be purified through acidic silica gel be injected into compound silica gel and purify in chromatographic column, with normal hexane and
Dichloromethane is according to 97:1～97:The pre- drip washing of the mixed solution of 4 volume ratio；It will be net chromatographic column will to be purified again through acidic silica gel
After change, concentrate eluant is fully transferred in compound silica gel purification chromatographic column；When liquid level is down to anhydrous sodium sulfate layer add just oneself
Alkane and dichloromethane are according to 97:1～97:The mixed solution of 4 volume ratio carries out eluting, eluent is concentrated into 0.5～
1.0mL；
The extracting solution for purifying chromatograph column purification through compound silica gel is injected into alkali alumina to purify in chromatographic column, will be through comprehensive silicon
After glue purifies chromatograph column purification, concentrate eluant is fully transferred in alkali alumina purification chromatographic column, when liquid level is down to anhydrous sulfur
Add normal hexane eluting alkali alumina to purify chromatographic column during sour sodium layer, dichloromethane is added when liquid level is down to anhydrous sodium sulfate layer
Alkane and the mixed solution eluting of normal hexane, then eluent is concentrated near dry；
(4), the concentrate eluant in step (3) is analyzed using gas chromatograph-mass spectrometer；
(5), quantitative analyses, using double internal standard quanitations, 8 kinds of Polychlorinated biphenyls Isotopic Internal Standards (13C12-labelled PCBs28、
52nd, 118,153,180,202,206 and 209) He 2 kind of Polychlorinated biphenyls isotope reclaim internal standard (13C12-PCB101、13C12-
PCB194) quantitative, Isotopic Internal Standard are used for the determinand for calibrating 33 kinds of Polychlorinated biphenyls.
10. according to claim 9 detection milk product in Polychlorinated biphenyls method, it is characterised in that：Gas phase in step (4)
The operating parameter of chromatograph-mass spectrometer is：The capillary chromatographic column of sampling (5%- phenyl)-methyl polysiloxane filler, resistance to height
The dottle pin of warm low-bleed and graphite grazing pad, deactivation are filled with glass cotton shunting and regardless of the universal bushing pipe of stream in processing；Column oven
Program be：Maintain 2 minutes at 100 DEG C first, then 180 DEG C are raised to the heating rate of 15 DEG C/min from 100 DEG C；Then with 3
DEG C/heating rate of min is raised to 240 DEG C from 180 DEG C；Finally 285 DEG C are raised to the heating rate of 10 DEG C/min from 240 DEG C, and
10min is maintained at 285 DEG C；Total run time is 42min；Carrier gas uses purity>99.999% high-purity helium, constant flow rate
For 1mL/min；The sample size of sample and standard liquid is 2.0 μ L, and using Splitless injecting samples pattern, injector temperature is 300 DEG C；Mass spectrum
Instrument parameter is：Ionization pattern, electron bombardment ionization source (EI), energy is 70eV；Ion source temperature is 230 DEG C, level Four bar temperature 150
DEG C, transmission line temperature is 300 DEG C, and solvent delay is 4min.
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| CN115524428B (en) * | 2022-01-07 | 2024-08-27 | 江苏微谱检测技术有限公司 | Method for measuring indicative polychlorinated biphenyl in food |
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