CN103331037A - Separation column filler for separating PCBs (Poly Chlorinated Biphenyls) or OCPs (Organic Chlorine Pesticides) in high-fat content sample and separation method - Google Patents
Separation column filler for separating PCBs (Poly Chlorinated Biphenyls) or OCPs (Organic Chlorine Pesticides) in high-fat content sample and separation method Download PDFInfo
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Abstract
The invention discloses a separation column filler for separating PCBs (Poly Chlorinated Biphenyls) or OCPs (Organic Chlorine Pesticides) in a high-fat content sample and a separation method. The filler is composed of 30%-35% of silica gel-florisil mixture, 50%-60% of acidic modified silica gel and 10%-15% of anhydrous sodium sulfate, by weight percentage. The separation column filler provided by the invention can be filled in a separation column for conventional column chromatography or a Seitz four-channel chromatographic separator, and used for chromatographic separation with the mixed solution of normal hexane and dichloromethane as a mobile phase. With the filler provided by the invention to separate the PCBs or the OCPs, the separation method is efficient, quick and simple in process; an obvious separation effect can be achieved under the condition of low cost; the column recovery rate of the PCBs materials is in the range from 93% to 105% and adding standard recovery to fish tissues is in the range from 74% to 100%; and the column recovery rate of the OCPs materials is in the range from 77% to 120% and adding standard recovery to fish tissues is in the range from 78% to 102%.
Description
Technical field
The present invention relates to a kind of filler of chromatography chromatography column, relate in particular to for separating of the separating column packing of PCBs or OCPs in the high fat content sample and utilize this filler to separate the method for PCBs in the fatty animal tissue sample or OCPs.
Background technology
Cleansing phase in the high fat content sample pretreatment process is mainly contained SPE (SPE), three kinds of common methods of gel permeation chromatography (GPC) and chromatography chromatography under the present stage technical conditions.
SPE (SPE) process can be divided into absorption and two parts of wash-out.In adsorption process, target substance optionally is adsorbed on has carried out enrichment on the adsorbent bed.Because the existence of factor such as coadsorption, adsorbent be selective, part also can be adsorbed at adsorbent bed the material that PCBs and OCPs have measured interference effect, influences separating effect in this process.SPE exists the individual components quantitative recovery incomplete in addition, analysis result is stable inadequately, the unsettled shortcoming of relative standard deviation [1] can not be used for incomplete volatile materials or nonvolatile matter that the extraction boiling point is higher than the solvent desorption temperature, uses shortcomings such as being subjected to certain limitation
[2]
Gel permeation chromatography (GPC), the core of GPC is that a chemically inert hollow bead is filled the pillar that forms, and when sample was flowed through filler mutually with flowing, macromolecular substances was owing to the micropore that can't the enter bead micromolecular weak point of stroke ratio that causes flowing through, prior to flowing out the last outflow of molecule minimum.GPC is particularly suitable for the removal of big molecule chaff interference such as lipid, albumen, nucleic acid in the complex environment sample (deposit and biological sample), separating effect for small-molecule substances such as PCBs, OCPs is general, and the equipment volume of GPC system is bigger, equipment cost is higher, use operating cost also higher, be difficult to promote on a large scale.
The chromatography chromatography, utilize that packing material adsorbs target substance to be measured in the chromatography chromatography column, remove impurity, then use eluent wash-out splitter, target substance wash-out to be measured is gone out, thereby realization isolation of purified from extract goes out the purpose of object to be measured.Because the amount of the adsorbent that splitter is filled is far longer than lamellae, with respect to SPE and GPC, the chromatography chromatogram can be used for the material of fractional dose bigger (grams magnitude).So the preparation as relatively large sample separates, its theoretical cam curve and post are imitated ratio far above the SPE post, and the chromatography chromatogram is better than SPE and GPC.In addition, chromatography chromatography equipment is simple, and flexibly changing eluant, eluent intensity obtains optimal separating effect, and this method be easy to the extension, with low cost, be to prepare the phosphatide graded product one of method of development potentiality is arranged most.The polarity of the particle diameter of column chromatography purification efficient and filler, specific area, the uniformity and mechanical performance and eluting solvent is relevant.Separation for the PCBs in animal tissue's sample or OCPs, though column chromatography has the rate of recovery and clean-up effect preferably, but the clean-up effect for the some of them composition is not ideal enough, as methoxychlor and the δ-BHC rate of recovery meeting variation that has certain polarity in organic chloro pesticide (OCPs), the conventional approach that improves is to strengthen the consumption of eluant, eluent, or suitably increase the polar intensity of eluant, eluent, caused solvent load big, shortcoming such as disengaging time is long
[3]For the high fat content sample, because of its matrix complexity, PCBs or OCP separate and need specially treated in the sample, and separation method is loaded down with trivial details, and disengaging time is long, and the rate of recovery is lower.Need exploitation that the most animals tissue sample is had good separating effect simultaneously, use quantity of solvent few, the isolation of purified method that separation cycle is short.
List of references:
[1] Yao Ziwei etc., organic micro-pollutant is analyzed pretreatment technology progress, marine environment and science,, the 3rd phase, 447-450 page or leaf in 2011 in the seawater.
[2] Cao Ling etc., the progress of the pretreatment technology that environmental organic pollutant detects, experimental technique and management,, the 3rd phase, 228-230 page or leaf in 2009.
[3] Dong Liang etc., organic pollutant analysis pre-treating method general introduction in the surrounding medium, Modern Scientific Instruments,, the 5th phase, 120-125 page or leaf in 2010.
Summary of the invention
The purpose of this invention is to provide a kind of chromatography chromatographic isolation column packing for separating of the PCBs in the high fat content sample or OCPs and utilize PCBs in this filler sample separation or the method for OCPs.Separating column packing of the present invention can be loaded in match thatch four-way chromatographic isolation instrument etc. and be separated into the chromatograph of principle with splitter, for flowing mutually, carries out the chromatography chromatographic isolation with the mixed solution of n-hexane and carrene.Separate, reclaim the sample solution gas chromatograph (GC-ECD) that obtains by the chromatography chromatography, gas chromatograph-mass spectrometer (GC-MS) etc. is analyzed.Utilize filler separating PCB s of the present invention or OCPs, separation method is efficient, quick, technology is simple, can be issued to significant separating effect in lower-cost condition, the PCBs material post rate of recovery can reach 93～105%, organizes recovery of standard addition can reach 74～100% to fish; The OCPs material post rate of recovery can reach 77～120%, organizes recovery of standard addition can reach 78～102% to fish.
Technical scheme of the present invention is:
A kind of separating column packing for separating of PCBs or OCPs is characterized in that, described filler is made up of silica gel-florisil silica (Florisil) mixture 30～35%, concentrated sulfuric acid modified silica-gel 50～60% and anhydrous sodium sulfate 10%～15% by mass percentage; The mass ratio of silica gel and florisil silica is 1:1 in wherein said silica gel-florisil silica mixture, the mass ratio of the concentrated sulfuric acid and silica gel is 45:55 in the described concentrated sulfuric acid modified silica-gel, and described filler is loaded in splitter successively by the order of silica gel-florisil silica mixture, concentrated sulfuric acid modified silica-gel, anhydrous sodium sulfate.
In above-mentioned filler, described silica gel, florisil silica (Florisil) and anhydrous sodium sulfate mix by proportioning after pre-treatment and activation respectively again or are used for modification.Described activation method will not limit, and can activate by the conventional activation method in this area.
The particle diameter of silica gel of the present invention is preferably 200～300 orders, and concentrated sulfuric acid modified silica-gel particle diameter is preferably 100～200 orders, and the florisil silica particle diameter is preferably 60～100 orders.
The present invention also provides a kind of splitter, and described splitter is fixing to be separating column packing of the present invention mutually.Separating column packing of the present invention is loaded the chromatography chromatographic isolation of carrying out PCBs or OCPs in splitter.When separation contains the sample solution of PCBs, use n-hexane: carrene=100:0～90:10 is separated for flowing, wherein the bed diameter/height of bed of splitter load filler is preferably 1:16, and sample solution contains at least a above PCBs that concentration respectively is 5～100ppb; When separation contains the OCPs sample solution, use n-hexane: carrene=25:75～15:85 is separated for flowing, wherein the bed diameter/height of bed of splitter load filler is preferably 1:16, and sample solution contains at least a above OCPs that concentration respectively is 5～100ppb.
Described splitter is preferably matched thatch four-way chromatographic isolation instrument splitter.Separating column packing of the present invention loaded in match thatch four-way chromatographic isolation instrument carry out the chromatography chromatographic isolation of PCBs or OCPs with splitter, wherein:
The method of separating PCB s is, carrying out column chromatography by match thatch four-way chromatographic isolation instrument separates, be specially: the sample solution that will contain PCBs is splined on the good described match thatch four-way chromatographic isolation instrument splitter of pre-balance, carries out wash-out with n-hexane: carrene=100:0～90:10 mutually for flowing; Wherein said sample solution contains at least a above PCBs that concentration respectively is 5～100ppb, the sample solution sample size is 1ml, bed diameter/the height of bed of described splitter load filler is 1:16, the phase wash-out pressure that flows is 70～80kPa, elution flow rate is 2～4mL/min, and the phase wash-out total amount that flows is 4～5 times of splitter retention volume.
The method of separating OCPs is, carrying out column chromatography by match thatch four-way chromatographic isolation instrument separates, be specially: the sample solution that will contain OCPs is splined on the good described match thatch four-way chromatographic isolation instrument splitter of pre-balance, carries out wash-out with n-hexane: carrene=25:75～15:85 mutually for flowing; Wherein said sample solution contains at least a above OCPs that concentration respectively is 5～100ppb, the sample solution sample size is 1ml, bed diameter/the height of bed of described splitter load filler is 1:16, flowing is n-hexane: carrene=25:75～15:85 mutually, the phase wash-out pressure that flows is 70～80kPa, elution flow rate is 2～4mL/min, and the phase wash-out total amount that flows is 5～6 times of splitter retention volume.
Match thatch four-way chromatographic isolation instrument of the present invention is the product of match thatch science and technology (Dalian) Co., Ltd, described match thatch four-way chromatographic isolation instrument splitter is match thatch four-way chromatographic isolation instrument dedicated separation post, comprise glass tube, the glass tube lower end is provided with sieve plate, and its concrete structure is referring to Chinese patent CN102419353B.
Beneficial effect of the present invention:
1. the present invention is silica gel, acid modification silica gel, and the mixture of florisil silica and anhydrous sodium sulfate is as the column chromatography for separation column packing, is applied to PCBs in the isolating environment medium or the separation of OCPs.Filler of the present invention is specially adapted to the separation of PCBs in the high fat content samples such as animal tissue or OCPs.Adopt the mixed fillers of different adsorption properties, the non-target substance in the high fat content sample is adsorbed on the separating column packing, make object see through splitter, realize the purpose that object separates with non-object (impurity).The corresponding eluent of statement in the invention that uses carries out wash-out to splitter, with object selectively wash-out go out.Simultaneously, acid modification silica gel can be eliminated the fatty carbonization in the sample and disturb, florisil silica adsorbs pigment in the sample, get rid of and disturb, realized the high selectivity isolation of purified of separating column packing to PCBs or OCPs in the higher fatty acid sample, reduced this stage of chaff interference wash-out has directly been carried out wash-out to object, improved separating effect and the rate of recovery.Experimental result shows, uses the filler of invention and the phase that flows, and the matrix recovery of standard addition of PCBs is 74～100% under the segregation ratio; The matrix recovery of standard addition of OCPs is 78～102%.
2. filler of the present invention, wherein silica gel, acid modification silica gel, the particle size range that florisil silica is selected is little, invents used packing material size, is reducing particle diameter, increase on the basis of specific area increase to non-object absorption property, consider to reduce the absorption to object simultaneously, make object can see through splitter, reach the purpose that object separates with impurity, realization is got rid of and is disturbed the removal of fat and pigment.
3. filler of the present invention is loaded in splitter, be the phase that flows with n-hexane-carrene, separating PCB s or OCPs, its separation method is simple, the solvent for use amount is few, and good separating effect is for traditional method rate of recovery low OCPs material such as methoxychlor, BHC has good separating effect, and its rate of recovery is all up to more than 75%.
4. filler of the present invention is loaded in match thatch four-way chromatographic isolation instrument splitter and is carried out the chromatography chromatographic isolation, for 21 kinds of PCBs or 18 kinds of OCPs in the animal tissue good separating effect and the rate of recovery are arranged all, its composition range has covered most of PCBs common in the present animal sample and OCPs material, and method of the present invention is applicable to separating and detection of PCBs and OCPs material in the multiple high fat content sample.
The specific embodiment
Following non-limiting example can make those of ordinary skill in the art more fully understand the present invention, but does not limit the present invention in any way.Test method described in the following embodiment if no special instructions, is conventional method; Described reagent and material if no special instructions, all can obtain from commercial channels, or can the conventional method preparation.
A kind of separating column packing for separating of PCBs or OCPs is characterized in that, described filler is made up of silica gel-florisil silica (Florisil) mixture 30～35%, concentrated sulfuric acid modified silica-gel 50～60% and anhydrous sodium sulfate 10%～15% by mass percentage; The mass ratio of silica gel and florisil silica is 1:1 in wherein said silica gel-florisil silica mixture, the mass ratio of the concentrated sulfuric acid and silica gel is 45:55 in the described concentrated sulfuric acid modified silica-gel, and described filler is loaded in splitter successively by the order of silica gel-florisil silica mixture, concentrated sulfuric acid modified silica-gel, anhydrous sodium sulfate.
In preferred embodiment, described filler is made up of silica gel-florisil silica mixture 35%, concentrated sulfuric acid modified silica-gel 55% and anhydrous sodium sulfate 10% by mass percentage;
The particle diameter of described silica gel is preferably the 200-300 order, and described concentrated sulfuric acid modified silica-gel particle diameter is preferably the 100-200 order, florisil silica particle diameter 60-100 order.
Described PCBs(polychlorinated biphenyls) is that biphenyl benzene ring hydrogen is replaced by chlorine and many chlorine compounds of forming, organism had a class persistence organic pollutant of savings property toxic action.
Described OCPs(organo-chlorine pesticide) be to prevent and treat the organic compound that contains the chlorine element in the constituent of phytopathy, insect pest.
Described filler is specially adapted to the separation of PCBs in the high fat content samples such as animal tissue or OCPs.
Above-mentioned filler is loaded the column chromatography for separation of carrying out PCBs in the sample or OCPs in splitter.When separation contains the sample solution of PCBs, use n-hexane: carrene=100:0～90:10 is separated for flowing, and the bed diameter/height of bed of splitter load filler is 1:16, and sample solution contains at least a above PCBs that concentration respectively is 5～100ppb; When separation contains the OCPs sample solution, use n-hexane: carrene=25:75～15:85 is separated for flowing, and the bed diameter/height of bed of splitter load filler is 1:16, and sample solution contains at least a above OCPs that concentration respectively is 5～100ppb.
Described splitter can be conventional column chromatography splitter, according to proportioning filler is loaded in splitter and sample solution on sample, under the natural gravity effect, carry out column chromatography for separation with the mixed solution of n-hexane and carrene.
Described splitter also can be separated into the employed splitter of chromatograph of principle for match thatch four-way chromatographic isolation instrument etc. with the chromatography chromatography column, according to proportioning filler is loaded in chromatographic isolation instrument splitter and sample solution on sample, be the mobile chromatographic isolation of carrying out mutually with the mixed solution of n-hexane and carrene.
The preliminary treatment of separating column packing:
Anhydrous sodium sulfate anhydrated down at 400 ℃ in dry 4 hours, and cooling is standby in the drier;
Florisil silica (particle diameter 60-100 order) anhydrated down at 500 ℃ in dry 12 hours, added the 6% deionized water activation of dry back florisil silica quality, left standstill more than the 6h after shaking up, and cooling is standby in the drier;
Silica gel (100-200 order and 200-300 order) down is cooled to normal temperature in the drier after dry 16 hours at 130 ℃, after 3.3% the deionized water that adds dry back silica gel gross weight fully shakes up, leaves standstill activation more than 6 hours.
Concentrated sulfuric acid modified silica-gel, activation back silica gel (100-200 order) and the concentrated sulfuric acid (98% analyze pure) according to quality than the concentrated sulfuric acid: silica gel=45:55 mixes.
Embodiment 1 match thatch four-way chromatographic isolation instrument separating PCB s mixture
The good separating column packing of preliminary treatment as stated above, load in match thatch four-way chromatographic isolation instrument (match thatch science and technology (Dalian) Co., Ltd) dedicated separation post, mixed solution with n-hexane and carrene separates the sample solution that contains 21 kinds of PCBs mutually for mobile, and concrete grammar is as follows:
(1) preparation of separating column packing
Take by weighing with good silica gel, concentrated sulfuric acid modified silica-gel, florisil silica and the anhydrous sodium sulfate of said method preliminary treatment, wherein the particle diameter of silica gel is the 200-300 order, and acid modification silica gel particle diameter is the 100-200 order, florisil silica particle diameter 60-100 order.Silica gel and the florisil silica mass ratio with 1:1 is mixed, stirs, obtain silica gel-florisil silica mixture.
(2) dress post
With 3g silica gel-florisil silica mixture, 5g concentrated sulfuric acid modified silica-gel, 1g anhydrous sodium sulfate, (the splitter internal diameter is 10mm to load match thatch four-way chromatographic isolation instrument splitter in vertical placement successively, length is 230mm, end is placed with sieve plate and prevents that filler from spilling) in, it forms lower floor, intermediate layer and upper strata respectively in splitter.Dry column-packing, dress are positioned over filler in the splitter naturally during post and get final product, and do not need compacting.Behind the dress post, the bed diameter/height of bed of load filler is 1:16.
(3) equilibrium separation post
Splitter upper end single adds 2ml n-hexane-carrene (100:0), wash-out under wash-out pressure 75kpa, and elution time is 47s, wash-out is 6-8 time repeatedly, makes splitter reach balance.
(4) separate
Slowly add PCBs that concentration is respectively 50ppb behind the column equilibration to be separated mixes mark solution and (contains 21 kinds of compositions just like table 1 to splitter upper end, U.S. Accustandard company Polychlorinated biphenyls mixes mark) 1ml, with n-hexane-methylene chloride volume than the mixed solution wash-out that is 100:0, wash-out pressure is 75kpa, each elution time is 70s, the eluent that at every turn adds 2ml is collected 8 eluents altogether and is amounted to 16ml in same collecting pipe; 16ml eluent rotary evaporation in 60 ℃ of water-baths is concentrated into about 1ml, dries up with nitrogen, add again behind the 1mL n-hexane dissolution and concentrate with nitrogen again, be settled to 1ml with n-hexane after 3 times repeatedly.
(5) measure
The sample solution that step (4) obtains is measured with gas chromatograph (GC-ECD), and calculated the rate of recovery that PCBs mixes each composition in the mark solution.
Result such as table 1.
Rate of recovery computational methods suc as formula:
The STD computational methods suc as formula:
In the following formula: STD is standard deviation; X
iBe the single experiment measured value; X is for repeatedly testing the mean value of getting; N is experiment number.
The RSD computational methods suc as formula:
Table 1.PCBs mixes in the mark solution 21 kinds of composition average recovery rates and relative standard deviation (%)
As known from Table 1, adopt the splitter of filler of the present invention to separate above 21 kinds of PCBs materials, can access good isolation of purified effect, the rate of recovery is between 93-105%, the RSD value meets US-EPA (Method3550,3500b less than 7%, 3500c, 3535a, 3600c, 3620b etc.) standard and Chinese national standard (GB/T9695.10-2008, GB/T5009.19-2008, GB/T9675-1998 etc.).The splitter stable in properties, reappearance is good, can be used for containing the isolation of purified of PCBs sample.
Embodiment 2 match thatch four-way chromatographic isolation instrument separate PCBs mixture in the fish tissue
(1) sample extraction
Take by weighing fish tissue sample (handling through freeze drying before the sample experiment) and be divided into experimental group 1 and experimental group 2, experimental group 1 is established 8 parallel tests, and experimental group 2 is established 4 parallel tests.Each experimental group adds the 5g fish respectively and is organized in the conical flask, and wherein adding concentration in the conical flask of experimental group 1 again is the mixed mark of the PCBs solution 1ml of 50ppb.Add acetone/benzinum (V in the conical flask of experimental group 1 and experimental group 2 respectively
1/ V
2=1/1) 20ml collects extract after 25 ℃ of ultrasonic waves extract 15min, extract is filtered by the funnel that is placed with anhydrous sodium sulfate, is collected in the conical flask.Use small amount of acetone/benzinum ((V again
1/ V
2=1/1) anhydrous sodium sulfate in the solution flushing funnel repeats said extracted process 3 times, merges all extracts.Extract rotary evaporation in 60 ℃ of water-baths is concentrated into about 1ml, gets testing sample.
(2) balance of splitter filling and splitter
Method operation according to (1) among the embodiment 1～(3).
(3) separate
The testing sample of each experimental group that obtains in the step (1) all is added into the splitter upper end after the balance respectively, with n-hexane-carrene (100:0) solvent elution, wash-out pressure is 75kpa, each elution time is 70s, the eluent that at every turn adds 2ml is collected 8 eluents altogether and is amounted to 16ml.16ml eluent rotary evaporation in 60 ℃ of water-baths is concentrated into about 1ml, dries up with nitrogen, concentrate with nitrogen again after adding the 1mL n-hexane dissolution, be settled to 1ml with n-hexane after 3 times repeatedly.
(4) measure
The sample solution that step (3) obtains is measured with gas chromatograph (GC-ECD), and calculated the rate of recovery that PCBs mixes each composition in the mark solution.
Result such as table 2.
Rate of recovery computational methods suc as formula:
The STD computational methods suc as formula:
Wherein in the following formula: STD is standard deviation; X
iBe the single experiment measured value; X is for repeatedly testing the mean value of getting; N is experiment number.
The RSD computational methods suc as formula:
21 kinds of PCBs average recovery rates and relative standard deviation (%) in the table 2. fish tissue
As known from Table 2, adopt the splitter of filler of the present invention to separate 21 kinds of PCBs materials in the above fish tissue, can access good isolation of purified effect, the rate of recovery is between 74-100%, the RSD value meets US-EPA (Method3550,3500b less than 15%, 3500c, 3535a, 3600c, 3620b etc.) standard and Chinese national standard (GB/T9695.10-2008, GB/T5009.19-2008, GB/T9675-1998 etc.).Splitter can play the isolation of purified effect to PCBs in the fish tissue, and the interference effect that can get rid of other materials (fat) in the fish tissue, realization is to the specific aim purification separation of PCBs material, reappearance is good, can be for the isolation of purified of high fat content sample PCBs such as fish tissue.
Embodiment 3 match thatch four-way chromatographic isolation instrument separate the OCPs mixture
(1) preparation of splitter and dress post
Method operation according to (1) among the embodiment 1～(2).
(2) equilibrium separation post
Splitter upper end single adds 2ml n-hexane-carrene (20:80), wash-out under wash-out pressure 75kpa, and elution time is 50s, wash-out is 7-9 time repeatedly, makes splitter reach balance.
(3) separate
Slowly add OCPs that concentration is respectively 50ppb behind the column equilibration to be separated mixes mark solution and (contains 22 kinds of OCPs compositions just like table 3 to splitter upper end, U.S. Accustandard company organochlorine (OCPs) mixes mark) 1ml, be the mixed solution wash-out of 20:80 with n-hexane-carrene ratio, wash-out pressure is 75kpa, each elution time is 76s, the eluent that at every turn adds 2ml is collected 9 eluents altogether and is amounted to 18ml in same tail type bottle; 18ml eluent rotary evaporation in 40 ℃ of water-baths is concentrated into about 1ml, dries up with nitrogen, concentrate with nitrogen again after adding the 1mL n-hexane dissolution, be settled to 1ml with n-hexane after 3 times repeatedly.
(4) measure
The sample solution that step (3) obtains is measured with gas chromatograph (GC-ECD), and calculated the rate of recovery that OCPs mixes each composition in the mark solution.
The computational methods of the rate of recovery are identical with embodiment 1.
Result such as table 3.
22 kinds of OCPs average recovery rates and relative standard deviation (%) in the table 3.OCPs solution
| Sequence number | The compound title | Average recovery rate | Relative standard deviation |
| 1 | hexachlorobenzene | 89 | 10 |
| 2 | α-HCH | 85 | 11 |
| 3 | γ-HCH | 98 | 8 |
| 4 | β-HCH | 81 | 4 |
| 5 | heptachlor | 102 | 8 |
| 6 | δ-HCH | 113 | 4 |
| 7 | Aldrin | - | - |
| 8 | Isodrin | - | - |
| 9 | heptachlor?epoxid | 104 | 6 |
| 10 | o,p′-DDE | 113 | 4 |
| 11 | Chlordane | 113 | 4 |
| 12 | endosulfan-1 | 77 | 7 |
| 13 | p,p′-DDE | 112 | 4 |
| 14 | Dieldrin | - | - |
| 15 | p,p′-DDD | 120 | 4 |
| 16 | endrin | - | - |
| 17 | O,p′-DDT | 119 | 8 |
| 18 | O,P′-DDD | 114 | 1 |
| 19 | endosulfan-2 | 77 | 8 |
| 20 | p,p′-DDT | 110 | 8 |
| 21 | Methoxychlor | 99 | 3 |
| 22 | Mirex | 118 | 5 |
As known from Table 3, adopt the splitter of filler of the present invention to separate above 22 kinds of OCPs materials, wherein 18 kinds of OCPs material rate of recovery are good, and Aldrin, Isodrin, Dieldrin and four kinds of materials of endrin are subjected to concentrated acid sulfonation effect loss serious, the rate of recovery is lower, can not realize separating purpose.Filler can play good isolation of purified effect to 18 kinds of materials wherein, the rate of recovery is between 77-120%, the RSD value meets US-EPA (Method3550,3500b less than 11%, 3500c, 3535a, 3600c, 3620b etc.) standard and Chinese national standard (GB/T9695.10-2008, GB/T5009.19-2008, GB/T9675-1998 etc.).The splitter stable in properties, reappearance is good, can be for the isolation of purified of high fat content sample OCPs such as fish tissue.
Embodiment 4 match thatch four-way chromatographic isolation instrument separate OCPs mixture in the fish tissue
(1) sample extraction
Take by weighing fish family tissue sample (freeze drying is handled before the experiment) and be divided into experimental group 1 and experimental group 2, experimental group 1 is established 8 parallel tests, and experimental group 2 is established 4 parallel tests.Each experimental group adds the 5g sample respectively in conical flask, and wherein adding concentration in the conical flask of experimental group 1 again is the mixed mark of the OCPs solution 1ml of 100ppb.Add acetone/benzinum (V in the conical flask of experimental group 1 and experimental group 2 respectively
1/ V
2=1/1) 20ml collects extract after 25 ℃ of ultrasonic waves extract 15min, extract is filtered by the funnel that is placed with anhydrous sodium sulfate, is collected in the triangular flask.Use small amount of acetone/benzinum (V again
1/ V
2=1/1) anhydrous sodium sulfate in the solution flushing funnel repeats said extracted process 3 times, merges all extracts.Extract rotary evaporation in 60 ℃ of water-baths is concentrated into about 1ml, gets testing sample.
(2) balance of splitter filling and splitter
Method operation according to (1) among the embodiment 3～(2).
(3) separate
The described testing sample of each experimental group all is added into the splitter upper end after the balance respectively, with n-hexane-carrene (20:80) solvent elution, wash-out pressure is 75kpa, and each elution time is 76s, the eluent that at every turn adds 2ml is collected 9 eluents altogether and is amounted to 18ml.18ml eluent rotary evaporation in 40 ℃ of water-baths is concentrated into about 1ml, dries up with nitrogen, concentrate with nitrogen again after adding the 1mL n-hexane dissolution, be settled to 1ml with n-hexane after 3 times repeatedly.
(4) measure
The sample solution that step (3) obtains is measured with gas chromatograph (GC-ECD), and calculated the rate of recovery that OCPs mixes each composition in the mark solution.
The computational methods of the rate of recovery are identical with embodiment 2, result such as table 4.
22 kinds of OCPs average recovery rates and relative standard deviation (%) in the table 4. fish tissue
As known from Table 4, adopt the splitter of filler of the present invention to separate above 22 kinds of OCPs materials, wherein 18 kinds of OCPs material rate of recovery are good, and Aldrin, Isodrin, Dieldrin and four kinds of materials of endrin are subjected to concentrated acid sulfonation effect loss serious, and the rate of recovery is lower.Filler can play good isolation of purified effect to 18 kinds of materials wherein, and the rate of recovery is between 78-102%, and the RSD value is less than 13%, meet US-EPA (Method3550,3500b, 3500c, 3535a, 3600c, 3620b etc.) standard and Chinese national standard (GB/T9695.10-2008, GB/T5009.19-2008, GB/T9675-1998 etc.), the splitter stable in properties, reappearance is good, can be for the isolation of purified of high fat content sample OCPs such as fish tissue.
Embodiment 5 column chromatography separating PCB s mixtures
(1) preparation of separating column packing
With reference to the method for (1) among the embodiment 1, prepare separating column packing.
(2) dress post
With 3g silica gel-florisil silica mixture, 5g concentrated sulfuric acid modified silica-gel, 1g anhydrous sodium sulfate, (the splitter internal diameter is 10mm to load match thatch four-way chromatographic isolation instrument splitter in vertical placement successively, length is 230mm, end is placed with sieve plate and prevents that filler from spilling) in, it forms lower floor, intermediate layer and upper strata respectively in splitter.Dry column-packing, dress are positioned over filler in the splitter naturally during post and get final product, and do not need compacting.Behind the dress post, the bed diameter/height of bed of load filler is 1:16.With the splitter vertical folder on iron stand.
(3) equilibrium separation post
Splitter upper end single adds 2ml n-hexane-carrene (100:0), wash-out under the natural gravity condition, and wash-out is 6-8 time repeatedly, makes splitter reach balance.
(4) separate
Slowly add the PCBs standard liquid that concentration is respectively 50ppb behind the column equilibration to be separated to splitter upper end and (contain 21 kinds of compositions just like table 5, U.S. Accustandard company Polychlorinated biphenyls mixes mark) 1ml, be the mixed solution wash-out of 100:0 with n-hexane-carrene ratio, the eluent that at every turn adds 2ml is collected 8 eluents altogether and is amounted to 16ml in same tail type bottle; 16ml eluent rotary evaporation in 60 ℃ of water-baths is concentrated into about 1-ml, dries up with nitrogen, add again behind the 1mL n-hexane dissolution and concentrate with nitrogen again, be settled to 1ml with n-hexane after 3 times repeatedly.
(5) measure
The sample solution that step (4) obtains is measured with gas chromatograph (GC-ECD), and calculated the rate of recovery that PCBs mixes each composition in the mark solution.
Computing formula is seen embodiment 1, result such as table 5
Table 5.PCBs mixes in the mark solution 21 kinds of composition average recovery rates and relative standard deviation (%)
As known from Table 5, adopt the splitter of filler of the present invention to separate above 21 kinds of PCBs materials, under the effect that breaks away from the chromatographic isolation instrument, still can access good isolation of purified effect, the rate of recovery is between 77-120%, and the RSD value meets US-EPA(Method3550 less than 8%, 3500b, 3500c, 3535a, 3600c, 3620b etc.) standard and Chinese national standard (GB/T9695.10-2008, GB/T5009.19-2008, GB/T9675-1998 etc.), reappearance is good.Filler still has the good rate of recovery under the condition of not using the chromatographic isolation instrument, can be applied under the non-separator state.Compare than the chromatographic isolation instrument, except the rate of recovery of PCB8 and PCB18 show slightly low, problem such as it is high that other composition rate of recovery show slightly, but exist systematic error bigger, and elution time is long.
Embodiment 6 column chromatographies separate PCBs mixture in the fish sample tissue
(1) sample extraction:
Method according to step (1) among the embodiment 2 is prepared testing sample.
(2) balance of splitter filling and splitter
Method operation according to (1) among the embodiment 5～(3).
(3) separate
The described testing sample of each experimental group that obtains in the step (1) all is added into the splitter upper end after the balance respectively, with n-hexane-carrene (100:0) solvent elution, adds the eluent of 2ml at every turn, collects 8 eluents altogether and amounts to 16ml.16ml eluent rotary evaporation in 60 ℃ of water-baths is concentrated into about 1ml, dries up with nitrogen, concentrate with nitrogen again after adding the 1mL n-hexane dissolution, be settled to 1ml with n-hexane after 3 times repeatedly.
(4) measure
The sample solution that step (3) obtains is measured with gas chromatograph (GC-ECD), and calculated the rate of recovery that PCBs mixes each composition in the mark solution.
Rate of recovery computing formula is seen embodiment 2, result such as table 6.
21 kinds of PCBs average recovery rates and relative standard deviation (%) in the table 6. fish tissue
| Sequence number | Compound | Average recovery rate | STD | Relative standard deviation |
| 1 | PCB8 | 87 | 0.07 | 8 |
| 2 | PCB18 | 81 | 0.04 | 5 |
| 3 | PCB28 | 82 | 0.06 | 7 |
| 4 | PCB52 | 84 | 0.06 | 7 |
| 5 | PCB44 | 85 | 0.06 | 7 |
| 6 | PCB77 | 84 | 0.06 | 7 |
| 7 | PCB101 | 86 | 0.07 | 8 |
| 8 | PCB66 | 83 | 0.07 | 8 |
| 9 | PCB126 | 89 | 0.06 | 7 |
| 10 | PCB153 | 97 | 0.07 | 8 |
| 11 | PCB118 | 88 | 0.06 | 7 |
| 12 | PCB138 | 94 | 0.08 | 9 |
| 13 | PCB105 | 92 | 0.06 | 6 |
| 14 | PCB187 | 90 | 0.06 | 7 |
| 15 | PCB128 | 90 | 0.06 | 6 |
| 16 | PCB180 | 98 | 0.06 | 6 |
| 17 | PCB170 | 95 | 0.06 | 6 |
| 18 | PCB195 | 94 | 0.06 | 6 |
| 19 | PCB200 | 99 | 0.06 | 6 |
| 20 | PCB206 | 104 | 0.06 | 6 |
| 21 | PCB209 | 101 | 0.05 | 5 |
As known from Table 6, invent described filler under the condition of not using the chromatographic isolation instrument, 21 kinds of PCBs rate of recovery are between 81%-104%, and RSD is less than 9%.Meet US-EPA(Method3550,3500b, 3500c, 3535a, 3600c, 3620b etc.) standard and Chinese national standard (GB/T9695.10-2008, GB/T5009.19-2008, GB/T9675-1998 etc.) requirement.Can realize the isolation of purified to PCBs in the fish tissue, can play catharsis to the impurity in the fish tissue (fat), especially can play the elimination effect to fat, reappearance is good, filler does not use under the situation of chromatographic isolation still can obtain the good rate of recovery, but than lower slightly under the instrument condition.Reappearance is stable, can be to the isolation of purified of PCBs in the high fat content samples such as fish tissue.
On the other hand: in embodiment 1～6, the mixed mark solution of PCBs or OCPs between its concentration 5～100ppb, all can access good isolation of purified effect, and the rate of recovery and RSD value all can reach described US-EPA standard and Chinese national standard.
Claims (8)
1. the separating column packing for separating of PCBs or OCPs is characterized in that, described filler is made up of silica gel-florisil silica mixture 30～35%, concentrated sulfuric acid modified silica-gel 50～60% and anhydrous sodium sulfate 10%～15% by mass percentage; The mass ratio of silica gel and florisil silica is 1:1 in wherein said silica gel-florisil silica mixture, the mass ratio of the concentrated sulfuric acid and silica gel is 45:55 in the described concentrated sulfuric acid modified silica-gel, and described filler is loaded in splitter successively by the order of silica gel-florisil silica mixture, concentrated sulfuric acid modified silica-gel, anhydrous sodium sulfate.
2. separating column packing according to claim 1 is characterized in that, the particle diameter of described silica gel is 200～300 orders, and concentrated sulfuric acid modified silica-gel particle diameter is 100～200 orders, and the florisil silica particle diameter is 60～100 orders.
3. a splitter is characterized in that, described splitter is fixing to be claim 1 or 2 described fillers mutually.
4. splitter according to claim 3 is characterized in that, described splitter is match thatch four-way chromatographic isolation instrument splitter.
5. the method for a separating PCB s is characterized in that, carries out column chromatography for separation by the described splitter of claim 3, and flowing is n-hexane: carrene=100:0～90:10 mutually.
6. the method for a separating PCB s, it is characterized in that, carrying out column chromatography by match thatch four-way chromatographic isolation instrument separates, be specially: the sample solution that will contain PCBs is splined on the described match of the good claim 4 of pre-balance thatch four-way chromatographic isolation instrument splitter, carries out wash-out with n-hexane: carrene=100:0～90:10 mutually for flowing; Wherein said sample solution contains at least a above PCBs that concentration respectively is 5～100ppb, the sample solution sample size is 1ml, bed diameter/the height of bed of described splitter load filler is 1:16, the phase wash-out pressure that flows is 70～80kPa, elution flow rate is 2～4mL/min, and the phase wash-out total amount that flows is 4～5 times of splitter retention volume.
7. a method of separating OCPs is characterized in that, carries out column chromatography for separation by the described splitter of claim 3, and flowing is n-hexane: carrene=25:75～15:85 mutually.
8. method of separating OCPs, it is characterized in that, carrying out column chromatography by match thatch four-way chromatographic isolation instrument separates, be specially: the sample solution that will contain OCPs is splined on the described match of the good claim 4 of pre-balance thatch four-way chromatographic isolation instrument splitter, carries out wash-out with n-hexane: carrene=25:75～15:85 mutually for flowing; Wherein said sample solution contains at least a above OCPs that concentration respectively is 5～100ppb, the sample solution sample size is 1ml, bed diameter/the height of bed of described splitter load filler is 1:16, flowing is n-hexane: carrene=25:75～15:85 mutually, the phase wash-out pressure that flows is 70～80kPa, elution flow rate is 2～4mL/min, and the phase wash-out total amount that flows is 5～6 times of splitter retention volume.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107638871A (en) * | 2017-09-30 | 2018-01-30 | 福建中烟工业有限责任公司 | Adsorbing composition and the method for determining dithio amino formate compounds |
| CN112444576A (en) * | 2020-11-04 | 2021-03-05 | 中山大学 | Method for improving detection precision of polychlorinated biphenyl in whale fish fat and application thereof |
| CN113945650A (en) * | 2021-09-09 | 2022-01-18 | 国家粮食和物资储备局科学研究院 | Method for simultaneously analyzing persistent organic pollutants in grains |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002228560A (en) * | 2001-02-01 | 2002-08-14 | Mitsubishi Heavy Ind Ltd | Detection device and method of organic halide |
| CN1563976A (en) * | 2004-03-24 | 2005-01-12 | 南京大学 | Synchronous purifying and extracting process for analysis of organic chlorine and organic phosphorus pesticide |
| JP2006313125A (en) * | 2005-05-09 | 2006-11-16 | Kansai Electric Power Co Inc:The | Method for speedily analyzing pcb on criterion of treated material of pcb waste |
| CN101871920A (en) * | 2010-06-13 | 2010-10-27 | 中国环境科学研究院 | Multistage improvement column for quickly pre-processing and purifying polychlorinated biphenyl in biological sample |
| CN102109505A (en) * | 2009-12-23 | 2011-06-29 | 中国科学院生态环境研究中心 | Pretreatment method for purifying extract liquor |
-
2013
- 2013-07-03 CN CN2013102754653A patent/CN103331037A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2002228560A (en) * | 2001-02-01 | 2002-08-14 | Mitsubishi Heavy Ind Ltd | Detection device and method of organic halide |
| CN1563976A (en) * | 2004-03-24 | 2005-01-12 | 南京大学 | Synchronous purifying and extracting process for analysis of organic chlorine and organic phosphorus pesticide |
| JP2006313125A (en) * | 2005-05-09 | 2006-11-16 | Kansai Electric Power Co Inc:The | Method for speedily analyzing pcb on criterion of treated material of pcb waste |
| CN102109505A (en) * | 2009-12-23 | 2011-06-29 | 中国科学院生态环境研究中心 | Pretreatment method for purifying extract liquor |
| CN101871920A (en) * | 2010-06-13 | 2010-10-27 | 中国环境科学研究院 | Multistage improvement column for quickly pre-processing and purifying polychlorinated biphenyl in biological sample |
Non-Patent Citations (2)
| Title |
|---|
| 赵荣彪，秦占芬，赵汝松，徐晓白: "水和生物样品中痕量多氯联苯的分析方法研究", 《分析化学研究报告》 * |
| 陈满英，张秀蓝，余乐洹，罗孝俊，麦碧娴: "沉积物中氯化石蜡与多氯联苯的分离及气相色谱-质谱检测", 《分析测试学报》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107638871A (en) * | 2017-09-30 | 2018-01-30 | 福建中烟工业有限责任公司 | Adsorbing composition and the method for determining dithio amino formate compounds |
| CN107638871B (en) * | 2017-09-30 | 2020-08-04 | 福建中烟工业有限责任公司 | Adsorption composition and method for determining dithiocarbamate compounds |
| CN112444576A (en) * | 2020-11-04 | 2021-03-05 | 中山大学 | Method for improving detection precision of polychlorinated biphenyl in whale fish fat and application thereof |
| CN112444576B (en) * | 2020-11-04 | 2021-12-28 | 中山大学 | Method for improving detection precision of polychlorinated biphenyl in whale fish fat and application thereof |
| CN113945650A (en) * | 2021-09-09 | 2022-01-18 | 国家粮食和物资储备局科学研究院 | Method for simultaneously analyzing persistent organic pollutants in grains |
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Application publication date: 20131002 |