CN104084180A - Method for preparing multilayer silica gel purification chromatographic column for detecting polybrominated biphenyls compounds - Google Patents
Method for preparing multilayer silica gel purification chromatographic column for detecting polybrominated biphenyls compounds Download PDFInfo
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Abstract
The invention provides a method for preparing and filling a multilayer silica gel purification chromatographic column. The method comprises the following steps: (1) preparing fillers, namely silver nitrate silica gel, sulfuric acid silica gel and sodium hydroxide silica gel, of the multilayer silica gel purification chromatographic column; (2) treating the fillers, namely silica gels and anhydrous sodium sulfate, of the multilayer silica gel purification chromatographic column; and (3) filling the multilayer silica gel purification chromatographic column. The multilayer silica gel purification chromatographic column prepared by adopting the method can overcome a complex matrix effect and adsorb and separate multiple impurities in a matrix containing polybrominated biphenyls compounds, so that the multiple impurities are effectively removed, and interference on subsequent detection of polybrominated biphenyls compounds is avoided. Moreover, the multilayer silica gel purification chromatographic column prepared by the method has the advantages of simplicity in operation, high reproducibility, high sensitivity and high accuracy.
Description
Technical field
The multilayer silica gel that the present invention relates to detect PBBs compounds purifies the preparation method of chromatographic column, belongs to environment or food inspection technical field.
Background technology
At present, organic pollutant category in environment and food is more and more, as PBBs compounds, and the residing matrix of this class organic pollution is also very complicated, in matrix, often contain the impurity such as various acid compounds, alkali compounds, aromatic compound, sulfur compound and aquo-compound, if these impurity are not treated, can produce larger interference to the testing result of follow-up organic pollution, the result that impact detects.Silica gel is as a kind of good absorption, although silica gel chromatographic column prepared by common silica gel chromatographic column preparation method can be removed the partial impurities in matrix, but it is more complicated to work as matrix situation, while detecting the polybrominated biphenyl compounds organic pollution in matrix, adopt silica gel chromatographic column prepared by common silica gel chromatographic column preparation method to be difficult to the Impurity removals such as various acid compounds, alkali compounds, aromatic compound, sulfur compound and aquo-compound in matrix, the interference that the detection of follow-up PBBs compounds is produced is still very large.Therefore the silica gel chromatographic column that, prepared by common silica gel chromatographic column preparation method can not be for detection of polybrominated biphenyl compounds organic pollution in complicated substrate.
Summary of the invention
The problem existing for solving prior art, the invention provides the preparation method of the multilayer silica gel purification chromatographic column that detects PBBs compounds, adopt multilayer silica gel purification chromatographic column prepared by the method can overcome complicated matrix effect, can be to carrying out adsorbing separation containing the plurality of impurities in the matrix of PBBs compounds, thereby effectively remove plurality of impurities, avoided the interference to follow-up polybrominated biphenyl compound test, and the multilayer silica gel prepared of the method purifies, and chromatographic column has advantages of simple to operate, favorable reproducibility, highly sensitive and the degree of accuracy is high.
Realizing the technical scheme that the object of the invention takes is:
The multilayer silica gel that detects PBBs compounds purifies a preparation method for chromatographic column, comprises the steps:
1.1 multilayer silica gel purify the preparation of chromatographic column filler silver nitrate silica gel, sulfuric acid silica gel and NaOH silica gel:
1.1.1 the preparation of silver nitrate silica gel:
The liquor argenti nitratis ophthalmicus that preparation mass percentage concentration is 10%～20%, liquor argenti nitratis ophthalmicus in mass ratio: silica gel=1:8～10, under stirring and lucifuge condition, liquor argenti nitratis ophthalmicus is added drop-wise in silica gel, liquor argenti nitratis ophthalmicus of every dropping drips next liquor argenti nitratis ophthalmicus after all will being stirred to and being uniformly dispersed again, after liquor argenti nitratis ophthalmicus dropwises, under lucifuge condition with oscillator by the mixed system sustained oscillation of liquor argenti nitratis ophthalmicus and silica gel to after completion of the reaction, under lucifuge condition, carry out stage drying, first at 50～70 ℃, toast 2～3 hours, then at 80～100 ℃, toast 2～3 hours, at 110～130 ℃, toast 4～5 hours again, obtain silver nitrate silica gel,
1.1.2 the preparation of sulfuric acid silica gel:
The concentrated sulfuric acid in mass ratio: silica gel=11:14～18, the dense sulfuric acid that is 95%～97% by mass percent concentration under stirring condition is added drop-wise in silica gel, concentrated sulfuric acid of every dropping drips next concentrated sulfuric acid after all will being stirred to and being uniformly dispersed again, after the concentrated sulfuric acid dropwises, with oscillator by the concentrated sulfuric acid with the mixed system sustained oscillation of silica gel to reacting complete, obtain sulfuric acid silica gel;
1.1.3 the preparation of NaOH silica gel:
Sodium hydroxide solution in mass ratio: silica gel=1:2～3, the sodium hydroxide solution that is 0.8mol/L～1.2mol/L by concentration under stirring condition is added drop-wise in silica gel, sodium hydroxide solution of every dropping drips next sodium hydroxide solution after all will being stirred to and being uniformly dispersed again, after dropwising, with oscillator by sodium hydroxide solution with the mixed system sustained oscillation of silica gel to reacting complete, obtain NaOH silica gel;
1.2 multilayer silica gel purify the processing of chromatographic column filler silica gel and anhydrous concentrated sulfuric acid sodium:
Silica gel was toasted after 10～12 hours at 240～260 ℃ standby, more anhydrous concentrated sulfuric acid sodium is toasted after 10～12 hours standby at 500～600 ℃;
1.3 multilayer silica gel purify the filling of chromatographic column:
1.3.1 in multilayer silica gel purification chromatographic column, fill order and the mass parts of each filler are from bottom to up:
1.3.2 get the chromatographic column of a dried and clean, described chromatographic column internal diameter and length ratio are 1:8～10, by mass parts described in step 1.3.1, take each filler, according to the fill order of step 1.3.1, each filler is poured into from the upper port of chromatographic column successively again, every kind of filler is from the upper port of chromatographic column pours into, allow the every kind of filler pouring into tile evenly, and chromatographic column inwall non-filler adheres to, during filling silver nitrate silica gel, silver nitrate silica gel filling good after, with masking foil, encase chromatographic column and make silver nitrate layer of silica gel lucifuge, all fillers are filling complete after, open the rotary-piston of chromatographic column lower end, lower port access vavuum pump in chromatographic column, open vavuum pump, while vacuumizing, with cork rod, beat the outer wall of chromatographic column, when the height of chromatographic column inner stuffing no longer declines, continue to vacuumize after at least 3 minutes, screw chromatographic column lower end piston,
1.3.3 open the piston of chromatographic column lower end, the flow velocity of liquid that makes to flow into chromatographic column is maximum, carrene is slowly injected to chromatographic column from chromatographic column upper port along chromatographic column inwall, make all fillers all wetted and without bubble, in chromatographic column, clear, the form of each filler level all for the moment, stop injecting carrene, finally use enough cyclohexane elution chromatography posts, the piston of closing chromatographic column lower end after wash-out.
Technique scheme is improved to step 1) in, while preparing silver nitrate silica gel, sulfuric acid silica gel and NaOH silica gel, be 6～8 hours with the time of oscillator concussion; Every kind of filler is from the upper port of chromatographic column pours into, and first, by the moving chromatographic column of have gentle hands jog, then with cork rod, the outer wall along chromatographic column beats gently, rotates chromatographic column, until chromatographic column inwall non-filler adheres to while beaing; During filling glass fibre, limit flattens the superficial compaction of glass layer with glass bar after beaing chromatographic column limit rotation chromatographic column to chromatographic column inwall and not having glass fibre to adhere to.
As shown from the above technical solution, the method has adopted different method of modifying to carry out modification to silica gel, silver nitrate silica gel, sulfuric acid silica gel and three kinds of modified silica-gels of NaOH silica gel have been obtained, silver nitrate silica gel can effectively adsorb containing the sulphur in the matrix of PBBs compounds and sulfur-containing compound, sulfuric acid silica gel can effectively adsorb containing the alkaline matter in the matrix of PBBs compounds, fat, protein and aromatic compound, and NaOH silica gel can effectively adsorb containing the acidic materials in the matrix of PBBs compounds.In the method for these three kinds of modified silica-gels of preparation, while dripping liquor argenti nitratis ophthalmicus, sulfuric acid and sodium hydroxide solution, rate of addition has strict demand, after every dropping liquor argenti nitratis ophthalmicus, the concentrated sulfuric acid or a sodium hydroxide solution, all must be stirred to and could drip next after fully mixing and drip, and after being added dropwise to complete, must shake 6～8 hours with oscillator.The advantages such as silver nitrate silica gel, sulfuric acid silica gel and the NaOH silica gel of adopting in this way preparation has good uniformity, decentralization is high and the ability of absorption impurity is strong.While preparing silver nitrate silica gel, adopt the method for stage drying to be dried silver nitrate silica gel, employing stage drying, temperature rising is mild, and silver nitrate silica gel is inside and outside all thoroughly and lentamente dry, has solved the problem that silver nitrate silica gel easily turns black and lumps.
In the method, at the bottom filled glass fiber of chromatographic column, glass fibre stable in properties, does not react with other compounds, in filling process, glass layer compacting is flattened, and can effectively support layer of silica gel and filter solid impurity.In the method, between each modified silicon glue-line, filling gel, as cushion, can effectively prevent that each modified silicon glue-line from reacting each other.Anhydrous sodium sulfate is filled by the superiors in chromatographic column in the method, can effectively remove the moisture in matrix.For the polybrominated biphenyl compounds organic pollution in environment and food, for the deimpurity interference that disappears, detect polybrominated biphenyl compounds organic pollution, in the method, the filler of chromatographic column has adopted special fill order and loading, this fill order and loading have following benefit: the sodium sulphate of the superiors is first removed the moisture in sample, silver nitrate silica gel is removed sulphur and sulfur-containing compound, sulphation silica gel is removed the alkali compounds in sample, fat, protein and aromatic compound, last NaOH SiClx glue is removed acid compound, can also neutralize the acid from sulphation layer of silica gel band.
The method adopts vacuum filling technology in filling process, makes filler even, fine and close, and filler surfacing, adds after solvent, and each filler can be all wetting by solvent, each filler form homogeneous, clear layer, and chromatogram matter is interior without bubble.
Compared with prior art, its beneficial effect and advantage are in the present invention:
The modified silica-gel superior performance of preparing in the method, and in the method, the fill order of filler is with strong points, fill order is reasonable, therefore, multilayer silica gel prepared by the method purifies chromatographic column can overcome complicated matrix effect, can remove containing the plurality of impurities in polybrominated biphenyl compounds organic pollution matrix, eliminate follow-up polybrominated biphenyl compounds organic pollution is detected to the interference producing.The more important thing is, it is simple to operate that multilayer silica gel prepared by the method purifies chromatographic column, the results showed, use multilayer silica gel chromatographic column prepared by the method detect the organic pollutions such as PBBs compounds have advantages of highly sensitive, the degree of accuracy is high, favorable reproducibility and the rate of recovery high.
The specific embodiment
The multilayer silica gel of preparing for the preparation method who checks the multilayer silica gel of detection PBBs compounds provided by the invention to purify chromatographic column purifies feasibility and the superiority of chromatographic column, the mixed standard solution that adds PBDE and metabolin thereof in water and bed mud, multilayer silica gel prepared by the preparation method who adopts the multilayer silica gel of detection PBBs compounds provided by the invention to purify chromatographic column the water of the mixed standard solution that contains PBDE and metabolin thereof and bed mud purifies chromatographic column and carries out adsorbing separation, the water of detection after multilayer silica gel purifies chromatographic column adsorbing separation and the content of the PBDE in bed mud and metabolin thereof, according to the result concentration detecting and known PBDE and the difference of metabolite concentration thereof, multilayer silica gel prepared by the method purifies chromatographic column feasibility and superiority is assessed.
Embodiment 1
1) prepare multilayer silica gel and purify chromatographic column
Take 10g silver nitrate, be dissolved in 100mL water, making mass percentage concentration is 10% liquor argenti nitratis ophthalmicus.Take 20g liquor argenti nitratis ophthalmicus, be dropwise added drop-wise in the silica gel 60 of 160g, one of every dropping all will ceaselessly be stirred to being uniformly dispersed with glass bar.After liquor argenti nitratis ophthalmicus dropwises, with oscillator, the mixed system of liquor argenti nitratis ophthalmicus and silica gel 60 was continued to shake after 8 hours, gains are carried out to stage drying processing, at 50 ℃, toast 3 hours; At 80 ℃, toast 3 hours; At 110 ℃, toast 5 hours, obtain silver nitrate silica gel, whole process is all wanted lucifuge, finally silver nitrate silica gel is stored in Brown Glass Brown glass bottles and jars only.
Take 55g mass percent concentration and be 97% the concentrated sulfuric acid, the concentrated sulfuric acid is dropwise added drop-wise in the silica gel 60 of 70g, one of every dropping all will ceaselessly be stirred after being uniformly dispersed and could drip next again with glass bar.After the concentrated sulfuric acid dropwises, with oscillator, by the mixed system concussion of sulfuric acid and silica gel 60 6 hours, obtain sulfuric acid silica gel, finally sulfuric acid silica gel is stored in Brown Glass Brown glass bottles and jars only.
Take 4g NaOH, be dissolved in 100mL water, be mixed with the sodium hydroxide solution that concentration is 1mol/L.Take 33g sodium hydroxide solution, be dropwise added drop-wise in the silica gel 60 of 66g, one of every dropping all will ceaselessly be stirred to being uniformly dispersed with glass bar.After sodium hydroxide solution dropwises, with oscillator, by the mixed system concussion of sodium hydroxide solution and silica gel 60 6 hours, obtain NaOH silica gel, finally NaOH silica gel is stored in Brown Glass Brown glass bottles and jars only.
Take 200g silica gel 60 and at 240 ℃, toast after 10 hours standbyly, then take 50g anhydrous sodium sulfate and at 500 ℃, toast after 12 hours standby.
Take 3 parts of quality and be respectively 2g, the silica gel 60 of 5g and 2g, take again 0.5g glass fibre, 3g NaOH silica gel, 12.5g sulfuric acid silica gel, 3g silver nitrate silica gel and 12.5g anhydrous sodium sulfate, get the chromatographic column of a dried and clean, the internal diameter of chromatographic column is 2cm, length is 19cm, according to glass fibre, silica gel 602g, NaOH silica gel, silica gel 605g, sulfuric acid silica gel, silica gel 602g, the order of silver nitrate silica gel and anhydrous sodium sulfate pours into each filler successively from the upper port of chromatographic column, every kind of filler is from the upper port of chromatographic column pours into, first by the moving chromatographic column of have gentle hands jog, allow the every kind of filler pouring into tile evenly, then with cork rod, the outer wall along chromatographic column beats gently, the rotation chromatographic column while beaing, the micro-silica gel that makes to be attached on chromatographic column inwall slips down, until chromatographic column inwall non-filler adheres to.During filling glass fibre, after limit is beaten chromatographic column limit rotation chromatographic column to chromatographic column inwall and is not had glass fibre to adhere to, with glass bar, the superficial compaction of glass layer is flattened, during filling silver nitrate silica gel, silver nitrate silica gel filling good after, with masking foil, encase chromatographic column and make silver nitrate layer of silica gel lucifuge.All fillers are filling complete after, open the rotary-piston of chromatographic column lower end, lower port access vavuum pump in chromatographic column, open vavuum pump, while vacuumizing, with cork rod, beat the outer wall of chromatographic column, when the height of chromatographic column inner stuffing no longer declines, continue to vacuumize 3 minutes, turn off vavuum pump and unload vavuum pump, screw chromatographic column lower end piston.Silica gel 60 used in the present embodiment can replace with other silica gel.
Open the piston of chromatographic column lower end to piston, the flow velocity of liquid that makes to flow into chromatographic column is maximum, carrene is slowly injected to chromatographic column from chromatographic column upper port along chromatographic column inwall, note not making filler be rushed by carrene, make filler whole when wetted and without bubble, in chromatographic column post, clear, the form of each filler level all for the moment, stops injecting carrene, finally use 50mL cyclohexane elution chromatography post, the piston of closing chromatographic column lower end after wash-out.
2) the multilayer silica gel of being prepared by said method purifies chromatographic column and assesses
Measure 500mL water sample, water sample is joined to 2L separatory funnel, add the mixed mark of 13 kinds of PBDEs solution (BDE17, BDE28, BDE47, BDE66, BDE71, BDE85, BDE99, BDE100, BDE138, BDE153, BDE154, BDE183, BDE190); The 8 kinds of mixed mark of methoxyl polybrominated diphenyl ethers solution (2 '-MeO-BDE68,6-MeO-BDE47,5-MeO-BDE47,4 '-MeO-BDE49,5 '-MeO-BDE100,4 '-MeO-BDE103,5 '-MeO-BDE99,4 '-MeO-BDE101); 7 kinds quantitatively with Isotopic Internal Standard Surrogate internal standards solution (
13c
12-BDE-28,
13c
12-BDE-47,
13c
12-BDE-99,
13c
12-BDE-100,
13c
12-BDE-153,
13c
12-BDE-154,
13c
12-BDE-183), 3 mark-on levels i.e. 0.05 μ g/L, 0.2 μ g/L, 0.5 μ g/L are set.Solution is mixed and balance 1～2h.
Add 60mL carrene to extract, after extraction, filter paper on folded on the separatory funnel through carrene rinse, on filter paper, pack the anhydrous sodium sulfate that volume is approximately funnel volume 1/2nd into, mixture after dichloromethane extraction is poured in funnel, and in the lower end of funnel, put the liquid that a concentrated bottle reception funnel flows out.
Repeat above-mentioned two steps twice, then clean separatory funnel with 20mL carrene, and in the lower end of separatory funnel, put the liquid that a concentrated bottle reception funnel flows out.Repeat the step 1 time that carrene cleans separatory funnel.
The liquid that each concentrated bottle graft is received mixes, and about 220mL, is then concentrated into 5mL and is transferred in teat glass; and add toluene or isooctane as protective agent; with Nitrogen evaporator, be concentrated near doing again, finally add the residue after 2mL n-hexane dissolution concentrates, obtain hexane extract.
Pour 2mL hexane extract into multilayer silica gel and purify in chromatographic column, with the n-hexane of 2mL, clean teat glass, altogether clean 2 times, and the liquid after cleaning is poured in multilayer silica gel purification chromatographic column.First with 50mL cyclohexane wash-out multilayer silica gel, purify chromatographic column; then with the mixing material wash-out multilayer silica gel of 40mL cyclohexane and 10mL carrene, purify chromatographic column; collect the liquid flowing out when twice wash-out multilayer silica gel purifies chromatographic column; the liquid that the chromatographic column that purifies multilayer silica gel flows out is receiving liquid; merge the receiving liquid of twice collection and be transferred in teat glass; add 2mL toluene or isooctane to make protective agent; with Nitrogen evaporator, be concentrated near doing again, finally add the residue after 1mL n-hexane dissolution concentrates.
With the content of PBBs compounds in the residue of isotope-dilution analysis or GC-MS method mensuration n-hexane dissolution, the result of mensuration is:
PBDE in water sample (13 kinds) and the rate of recovery of methoxyl polybrominated diphenyl ethers (8 kinds) under the mark-on level of 0.05 μ g/L are: 72%～90%;
PBDE in water sample (13 kinds) and the rate of recovery of methoxyl polybrominated diphenyl ethers (8 kinds) under the mark-on level of 0.2 μ g/L are: 78%～103%;
PBDE in water sample (13 kinds) and the rate of recovery of methoxyl polybrominated diphenyl ethers (8 kinds) under the mark-on level of 0.5 μ g/L are: 86%～102%.
The result of the above-mentioned rate of recovery meets the related request in european union directive (European Union document2002/657/EC).While being P < 1 μ g/L, the rate of recovery need be in 50%～120% scope, and P refers to the concentration of determinand.
Embodiment 2
1) prepare multilayer silica gel and purify chromatographic column
Take 20g silver nitrate, be dissolved in 100mL water, making mass percentage concentration is 20% liquor argenti nitratis ophthalmicus.Take 20g liquor argenti nitratis ophthalmicus, be dropwise added drop-wise in the silica gel 60 of 200g, one of every dropping all will ceaselessly be stirred after being uniformly dispersed and could drip next with glass bar.After liquor argenti nitratis ophthalmicus dropwises, with oscillator, the mixed system of liquor argenti nitratis ophthalmicus and silica gel 60 was continued to shake after 6 hours, gains are carried out to stage drying processing, at 70 ℃, toast 2 hours; At 100 ℃, toast 2 hours; At 130 ℃, toast 4 hours, obtain silver nitrate silica gel, whole process is all wanted lucifuge, finally silver nitrate silica gel is stored in Brown Glass Brown glass bottles and jars only.
The concentrated sulfuric acid that the concentration that takes 55g is 95%, is dropwise added drop-wise to the concentrated sulfuric acid in the silica gel 60 of 90g, and one of every dropping all will ceaselessly be stirred to being uniformly dispersed with glass bar.After the concentrated sulfuric acid dropwises, with oscillator, by the mixed system concussion of the concentrated sulfuric acid and silica gel 60 8 hours, obtain sulfuric acid silica gel, finally sulfuric acid silica gel is stored in Brown Glass Brown glass bottles and jars only.
Take 4.8g NaOH, be dissolved in 100mL water, be mixed with the sodium hydroxide solution that concentration is 1.2mol/L.Take 33g sodium hydroxide solution, be dropwise added drop-wise in the silica gel 60 of 99g, one of every dropping all will ceaselessly be stirred to being uniformly dispersed with glass bar.After sodium hydroxide solution dropwises, with oscillator, by the mixed system concussion of sodium hydroxide solution and silica gel 60 8 hours, obtain NaOH silica gel, finally NaOH silica gel is stored in Brown Glass Brown glass bottles and jars only.
Take 200g silica gel 60 and at 260 ℃, toast after 12 hours standbyly, then take 50g anhydrous sodium sulfate and at 550 ℃, toast after 12 hours standby.
Take 3 parts of quality and be respectively 3g, the silica gel 60 of 7.5g and 3g, take again 1g glass fibre, 4.5g NaOH silica gel, 15g sulfuric acid silica gel, 4.5g silver nitrate silica gel and 15g anhydrous sodium sulfate, get the chromatographic column of a dried and clean, the internal diameter of chromatographic column is 2.5cm, length is 25cm, according to glass fibre, silica gel 603g, NaOH silica gel, silica gel 607.5g, sulfuric acid silica gel, silica gel 603g, the order of silver nitrate silica gel and anhydrous sodium sulfate pours into each filler successively from the upper port of chromatographic column, every kind of filler is from the upper port of chromatographic column pours into, first by the moving chromatographic column of have gentle hands jog, allow each filler pouring into tile evenly, then with cork rod, the outer wall along chromatographic column beats gently, the rotation chromatographic column while beaing, the micro-silica gel that makes to be attached on chromatographic column inwall slips down, until chromatographic column inwall non-filler adheres to.During filling glass fibre, limit flattens the superficial compaction of glass layer with glass bar after beaing chromatographic column limit rotation chromatographic column to chromatographic column inwall and not having glass fibre to adhere to.During filling silver nitrate silica gel, silver nitrate silica gel filling good after, with masking foil, encase chromatographic column and make silver nitrate layer of silica gel lucifuge.All fillers are filling complete after, open the rotary-piston of chromatographic column lower end, lower port access vavuum pump in chromatographic column, open vavuum pump, while vacuumizing, with cork rod, beat the outer wall of chromatographic column, when the height of chromatographic column inner stuffing no longer declines, continue to vacuumize 5 minutes, turn off vavuum pump and unload vavuum pump, screw chromatographic column lower end piston.
Open the piston of chromatographic column lower end to piston, the flow velocity of liquid that makes to flow into chromatographic column is maximum, carrene is slowly injected to chromatographic column from chromatographic column upper port along chromatographic column inwall, note not making filler be rushed by carrene, make filler whole when wetted and without bubble, in chromatographic column post, clear, the form of each filler level all for the moment, stops injecting carrene, finally use 80mL cyclohexane elution chromatography post, the piston of closing chromatographic column lower end after wash-out.
2) the multilayer silica gel of being prepared by said method purifies chromatographic column and assesses
Taking 10g bed mud sample joins in mill, add 3g anhydrous sodium sulfate (adding anhydrous sodium sulfate is in order to absorb the moisture in sample), then add the mixed mark of 13 kinds of PBDEs solution (BDE17, BDE28, BDE47, BDE66, BDE71, BDE85, BDE99, BDE100, BDE138, BDE153, BDE154, BDE183, BDE190); The 8 kinds of mixed mark of methoxyl polybrominated diphenyl ethers solution (2 '-MeO-BDE68,6-MeO-BDE47,5-MeO-BDE47,4 '-MeO-BDE49,5 '-MeO-BDE100,4 '-MeO-BDE103,5 '-MeO-BDE99,4 '-MeO-BDE101); 7 kinds quantitatively with Isotopic Internal Standard Surrogate internal standards solution (
13c
12-BDE-28,
13c
12-BDE-47,
13c
12-BDE-99,
13c
12-BDE-100,
13c
12-BDE-153,
13c
12-BDE-154,
13c
12-BDE-183), 3 mark-on levels i.e. 2.5 μ g/kg, 10 μ g/kg, 25 μ g/kg are set.Solution is mixed and balance 1～2h, transfer in filtration paper cylinder.
Add the mixing material of 125mL acetone and 125mL carrene, then adopt the method that Soxhlet is extracted to extract 16 hours, and collect the extract after extracting, the about 250mL of extract.Extract is concentrated into after 5～10mL, is transferred in teat glass, add 2mL toluene or isooctane to make protective agent, then be concentrated near doing with Nitrogen evaporator, finally add the residue after 2mL n-hexane dissolution concentrates, obtain hexane extract.
Pour 2mL hexane extract into multilayer silica gel and purify in chromatographic column, with the n-hexane of 2mL, clean teat glass, altogether clean 2 times, and the liquid after cleaning is poured in multilayer silica gel purification chromatographic column.First with 50mL cyclohexane wash-out multilayer silica gel, purify chromatographic column; then with the mixing material wash-out multilayer silica gel of 40mL cyclohexane and 10mL carrene, purify chromatographic column; collect the liquid flowing out when twice wash-out multilayer silica gel purifies chromatographic column; the liquid that the chromatographic column that purifies multilayer silica gel flows out is receiving liquid; merge the receiving liquid of twice collection and be transferred in teat glass; add 2mL toluene or isooctane to make protective agent; with Nitrogen evaporator, be concentrated near doing again, finally add the residue after 1mL n-hexane dissolution concentrates.
With the content of PBBs compounds in the residue of isotope-dilution analysis or GC-MS method mensuration n-hexane dissolution, the result of mensuration is:
PBDE in bed mud (13 kinds) and the rate of recovery of methoxyl polybrominated diphenyl ethers (8 kinds) under the mark-on level of 2.5 μ g/kg are: 72%～92%;
PBDE in bed mud (13 kinds) and the rate of recovery of methoxyl polybrominated diphenyl ethers (8 kinds) under the mark-on level of 10 μ g/kg are: 84%～103%;
PBDE in bed mud (13 kinds) and the rate of recovery of methoxyl polybrominated diphenyl ethers (8 kinds) under the mark-on level of 25 μ g/kg are: 93%～100%.
The result of the above-mentioned rate of recovery meets the related request in european union directive (European Union document2002/657/EC).The rate of recovery need be in 70%～110% scope during 1 μ g/kg≤P < 10 μ g/kg; During 10 μ g/kg≤P < 100 μ g/kg, the rate of recovery need be in 80%～110% scope, and P refers to the concentration of determinand.
According to the result of embodiment 1 and embodiment 2 detections, can find out, the rate of recovery of each persistent organism all meets the related request in european union directive (European Union document2002/657/EC), illustrate adopt multilayer silica gel provided by the invention to purify the preparation of chromatographic column and multilayer silica gel chromatographic column prepared by fill method can be for detection of the content of persistent organism in environmental sample, and highly sensitive, the degree of accuracy is high, favorable reproducibility and the rate of recovery high.
Claims (4)
1. the multilayer silica gel that detects PBBs compounds purifies the preparation method of chromatographic column, it is characterized in that comprising the steps:
1.1 multilayer silica gel purify the preparation of chromatographic column filler silver nitrate silica gel, sulfuric acid silica gel and NaOH silica gel:
1.1.1 the preparation of silver nitrate silica gel:
The liquor argenti nitratis ophthalmicus that preparation mass percentage concentration is 10%～20%, liquor argenti nitratis ophthalmicus in mass ratio: silica gel=1:8～10, under stirring and lucifuge condition, liquor argenti nitratis ophthalmicus is added drop-wise in silica gel, liquor argenti nitratis ophthalmicus of every dropping drips next liquor argenti nitratis ophthalmicus after all will being stirred to and being uniformly dispersed again, after liquor argenti nitratis ophthalmicus dropwises, under lucifuge condition with oscillator by the mixed system sustained oscillation of liquor argenti nitratis ophthalmicus and silica gel to after completion of the reaction, under lucifuge condition, carry out stage drying, first at 50～70 ℃, toast 2～3 hours, then at 80～100 ℃, toast 2～3 hours, at 110～130 ℃, toast 4～5 hours again, obtain silver nitrate silica gel,
1.1.2 the preparation of sulfuric acid silica gel:
The concentrated sulfuric acid in mass ratio: silica gel=11:14～18, the dense sulfuric acid that is 95%～97% by mass percent concentration under stirring condition is added drop-wise in silica gel, concentrated sulfuric acid of every dropping drips next concentrated sulfuric acid after all will being stirred to and being uniformly dispersed again, after the concentrated sulfuric acid dropwises, with oscillator by the concentrated sulfuric acid with the mixed system sustained oscillation of silica gel to reacting complete, obtain sulfuric acid silica gel;
1.1.3 the preparation of NaOH silica gel:
Sodium hydroxide solution in mass ratio: silica gel=1:2～3, the sodium hydroxide solution that is 0.8mol/L～1.2mol/L by concentration under stirring condition is added drop-wise in silica gel, sodium hydroxide solution of every dropping drips next sodium hydroxide solution after all will being stirred to and being uniformly dispersed again, after dropwising, with oscillator by sodium hydroxide solution with the mixed system sustained oscillation of silica gel to reacting complete, obtain NaOH silica gel;
1.2 multilayer silica gel purify the processing of chromatographic column filler silica gel and anhydrous concentrated sulfuric acid sodium:
Silica gel was toasted after 10～12 hours at 240～260 ℃ standby, more anhydrous concentrated sulfuric acid sodium is toasted after 10～12 hours standby at 500～600 ℃;
1.3 multilayer silica gel purify the filling of chromatographic column:
1.3.1 in multilayer silica gel purification chromatographic column, fill order and the mass parts of each filler are from bottom to up:
1.3.2 get the chromatographic column of a dried and clean, described chromatographic column internal diameter and length ratio are 1:8～10, by mass parts described in step 1.3.1, take each filler, according to the fill order of step 1.3.1, each filler is poured into from the upper port of chromatographic column successively again, every kind of filler is from the upper port of chromatographic column pours into, allow the every kind of filler pouring into tile evenly, and chromatographic column inwall non-filler adheres to, during filling silver nitrate silica gel, silver nitrate silica gel filling good after, with masking foil, encase chromatographic column and make silver nitrate layer of silica gel lucifuge, all fillers are filling complete after, open the rotary-piston of chromatographic column lower end, lower port access vavuum pump in chromatographic column, open vavuum pump, while vacuumizing, with cork rod, beat the outer wall of chromatographic column, when the height of chromatographic column inner stuffing no longer declines, continue to vacuumize after at least 3 minutes, screw chromatographic column lower end piston,
1.3.3 open the piston of chromatographic column lower end, the flow velocity of liquid that makes to flow into chromatographic column is maximum, carrene is slowly injected to chromatographic column from chromatographic column upper port along chromatographic column inwall, make all fillers all wetted and without bubble, in chromatographic column, clear, the form of each filler level all for the moment, stop injecting carrene, finally use enough cyclohexane elution chromatography posts, the piston of closing chromatographic column lower end after wash-out.
2. the multilayer silica gel that detects according to claim 1 PBBs compounds purifies the preparation method of chromatographic column, it is characterized in that: step 1) in, while preparing silver nitrate silica gel, sulfuric acid silica gel and NaOH silica gel, be 6～8 hours with the time of oscillator concussion.
3. the multilayer silica gel that detects according to claim 1 PBBs compounds purifies the preparation method of chromatographic column, it is characterized in that: every kind of filler is from the upper port of chromatographic column pours into, first by the moving chromatographic column of have gentle hands jog, then with cork rod, the outer wall along chromatographic column beats gently, the rotation chromatographic column while beaing, until chromatographic column inwall non-filler adheres to.
4. the multilayer silica gel that detects according to claim 1 PBBs compounds purifies the preparation method of chromatographic column, it is characterized in that: during filling glass fibre, limit flattens the superficial compaction of glass layer with glass bar after beaing chromatographic column limit rotation chromatographic column to chromatographic column inwall and not having glass fibre to adhere to.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4658000A (en) * | 1983-08-16 | 1987-04-14 | Reanal Finomvegyszergyar | Polyacrylamide adhesive for fixing the sorbent layers of overpressured, one-and multilayer-chromatographic plates and a process for the preparation thereof |
| CN101652651A (en) * | 2007-03-29 | 2010-02-17 | 国立大学法人爱媛大学 | Method of extracting polychlorinated biphenyl |
| CN102463103A (en) * | 2010-11-18 | 2012-05-23 | 甘肃省分析测试中心 | Method for preparing ethanediamine-N-propoxysilane solid-phase column extractor filling materials |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4658000A (en) * | 1983-08-16 | 1987-04-14 | Reanal Finomvegyszergyar | Polyacrylamide adhesive for fixing the sorbent layers of overpressured, one-and multilayer-chromatographic plates and a process for the preparation thereof |
| CN101652651A (en) * | 2007-03-29 | 2010-02-17 | 国立大学法人爱媛大学 | Method of extracting polychlorinated biphenyl |
| CN102463103A (en) * | 2010-11-18 | 2012-05-23 | 甘肃省分析测试中心 | Method for preparing ethanediamine-N-propoxysilane solid-phase column extractor filling materials |
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