CN111175400A - Organic matter enrichment and concentration method based on low-pollution water body chemical and biological analysis - Google Patents
Organic matter enrichment and concentration method based on low-pollution water body chemical and biological analysis Download PDFInfo
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- CN111175400A CN111175400A CN202010030090.4A CN202010030090A CN111175400A CN 111175400 A CN111175400 A CN 111175400A CN 202010030090 A CN202010030090 A CN 202010030090A CN 111175400 A CN111175400 A CN 111175400A
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Abstract
The invention relates to an organic matter enrichment and concentration method based on low-pollution water body chemistry and biological analysis, which optimizes the retention condition of organic matters in sewage in a subsequent solid-phase extraction cartridge by acidifying a sewage water sample and adding an organic solvent methanol into the water sample; optimizing the adsorption condition and subsequent elution condition of the solid-phase extraction column on the organic matters in the sewage by adjusting the type and volume ratio of the organic solvent; and optimizing the dissolving condition of the obtained concentrated organic matters by adjusting the solvent of the organic matters. The method can obtain the total organic matter recovery rate of the sewage of more than 85 percent, and the concentrated organic matter is enriched and can be used for the quantitative analysis of the subsequent trace organic matters and the qualitative analysis of the biological effect. The method comprehensively considers the recovery of small molecular organic matters and large molecular organic matters in the sewage, has high accuracy and good recovery rate, and provides a scientific and effective water sample pretreatment method for monitoring trace organic pollutants and biotoxicity effect in the sewage.
Description
Technical Field
The invention relates to the technical field of detection of environmental water, in particular to an organic matter enrichment and concentration method based on chemical and biological analysis of low-pollution water.
Background
Soluble organic matters in secondary effluent of a sewage plant account for more than 90 percent of total organic matters, and the substances are various in types and complicated in nature, namely trace organic pollutant containing micromolecules, macromolecular polysaccharide, protein, humic acid and other substances. At present, trace organic pollutants detected in secondary effluent of a sewage plant comprise Endocrine Disruptors (EDCs), medicaments and personal care products (PPCPs), Organic Phosphorus Flame Retardants (OPFRs), pesticide insecticides and the like, and the substances can generate a bioaccumulation effect in a sewage receiving water body and can directly generate toxicity to aquatic organisms; adverse biological effects detected in sewage plant secondary effluent include acute toxicity of non-specific luminescent bacteria, specific endocrine disrupting effects, active genotoxic umuC genotoxicity, and phytophotosynthetic inhibitory effects. With the concern of people on the ecological safety of water and the development of detection technology, the quantitative detection of trace organic pollutants combined with a biotoxicity effect detection means has become an inevitable trend in the field of sewage monitoring in the future. However, efficient and accurate detection requires scientific and efficient water sample pretreatment steps. At present, the pretreatment of a water sample only considers the recovery of trace organic pollutants, but does not consider the recovery of macromolecular organic matters; or only the recovery of macromolecular organic matters is considered, and the effective recovery rate cannot be ensured. Therefore, it is not favorable for the subsequent biological toxicity detection and the qualitative and quantitative analysis of the physicochemical indexes of the organic matters. The scientific and reasonable water sample pretreatment method ensures the recovery of small molecular trace organic pollutants in sewage and macromolecular organic matters in the sewage on one hand, and maximizes the recovery rate of each component in the sewage so as to ensure the accuracy of subsequent detection.
Disclosure of Invention
In order to solve the technical problems, the invention aims to provide an organic matter enrichment and concentration method based on chemical and biological analysis of a low-pollution water body, comprehensively considers the recovery of trace organic pollutants, macromolecular polysaccharides, proteins, humus and the like in sewage, and improves the recovery rate of target organic matters, thereby ensuring the accuracy of subsequent biotoxicity detection and physicochemical index analysis.
In order to achieve the purpose, the invention adopts the technical scheme that:
the organic matter enriching and concentrating method based on the chemical and biological analysis of the low-pollution water body comprises the following steps:
step one, pretreating a sewage sample:
collecting a sewage water sample according to a standard sampling method, and treating the water sample as follows:
(1) filtering a certain amount of sewage water sample by a glass fiber filter membrane by using a large-volume filtering device;
(2) storing the filtered water sample in a glass container by using H2SO4Acidifying the pH of the water sample to 2.5;
(3) adding a certain amount of chromatographic grade methanol solution into a water sample to ensure that the final volume fraction is 5%;
step two, enriching and concentrating organic matters:
utilizing a solid phase extraction device to enrich and concentrate organic matters in a water sample:
(1) connecting a C18 solid phase extraction column and an HLB solid phase extraction column in series;
(2) sequentially passing 10mL of dichloromethane and n-hexane mixed solution (volume ratio is 1:1), 10mL of methanol and 10mL of ultrapure water (pH is 2.5) through a solid phase extraction column connected in series for activation and cleaning;
(3) continuously passing the water sample obtained in the first step in the glass container through the serially connected solid phase extraction small columns, and adjusting the pressure of a vacuum pump;
(4) after the sample loading of the sewage water sample is finished, cleaning the tandem small column by using 10mL of ultrapure water (pH 2.5);
(5) continuously performing vacuum pumping by using a solid phase extraction device to fully dry the small columns connected in series, and then putting the small columns into a centrifugal machine for further dehydration and drying;
step three, preparing a concentrated organic solution:
(1) sequentially passing 10mL of methanol, 10mL of dichloromethane and n-hexane mixed solution (volume ratio is 1:1) through the dried solid phase extraction small column, and collecting eluent;
(2) blowing the eluent to about 2mL by using high-purity nitrogen under the water bath condition, then concentrating all the eluent into a glass test tube, and blowing the eluent to a fully dry state by using high-purity nitrogen;
(3) dissolving the obtained solid organic matters by using phosphate solution, wherein the final concentration multiple of the organic matters is determined according to the biological toxicity detection result (calculated by TOC);
(4) and dissolving the obtained solid organic matters by using an organic solvent, and using the dissolved solid organic matters for subsequent sample loading detection of a liquid chromatography-mass spectrometer or a gas chromatography-mass spectrometer.
In the first step, the water sample pretreatment is specifically combined with sewage water sample acidification (pH 2.5) and 5% methanol organic solvent addition (volume fraction).
In the step one, the glass fiber filter membrane is 0.7/0.45 mu m, and H2SO4The concentration was 2M.
In the second step, the mixture of dichloromethane and n-hexane (volume ratio of 1:1) and methanol are used in a ratio of 1:1, and the polarity coverage of the activated solvent is 0.06 (n-hexane) → 3.4 (dichloromethane) → 6.6 (methanol).
And in the second step, the water sample overflowing speed is kept below 5mL/min, and the solid phase extraction device is used for continuously vacuumizing for 20 min.
In the third step, the mixture of dichloromethane and n-hexane (volume ratio is 1:1) and methanol are used in proportion (volume ratio is 1: 1).
The water bath condition in the third step is 40 ℃, and the phosphate solution is 5mM (pH 8).
The invention has the beneficial effects that:
①, in the invention, the recovery rate of the sewage organic matter can reach more than 85 percent while the desalination is realized in the process of enriching and concentrating the organic matter, and the optical property of the enriched and concentrated organic matter is basically consistent with that of the raw water organic matter;
in the invention, the obtained organic matter concentrated solution can be directly used for detecting trace organic pollutants of a liquid chromatography-mass spectrometer and a gas chromatography-mass spectrometer and can also be used for detecting biological toxicity;
the invention greatly reduces the volume of water sample for sewage organic matter quantitative detection and biological effect qualitative detection, and improves the sensitivity of analysis and detection.
Drawings
FIG. 1 is a schematic flow chart of a design method of the present invention.
The TOC and UV-vis index of organic material obtained by the method of the present invention and the prior art method in the example of FIG. 2 were analyzed.
FIG. 3 illustrates the analysis of three-dimensional fluorescence spectrum indicators of organic materials obtained by the method of the present invention and the prior art.
Fig. 4 illustrates the analysis of trace organic contaminants in organic materials obtained by the method of the present invention and the prior art method.
FIG. 5 is a graph showing the analysis of acute toxicity of organic luminescent bacteria obtained by the method of the present invention and the method of the prior art.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
Example 1:
as shown in fig. 1: collecting a 20L water sample of effluent of a secondary sedimentation tank from a typical sewage treatment plant according to a standard method, wherein 10L water sample is carefully subjected to sample treatment according to the method; another 10L of water sample was carefully sample treated according to the usual methanol method: filtering a certain amount of sewage water sample through a 0.7-micron glass fiber filter membrane; activating and cleaning the HLB column by using 20mL of methanol and 10mL of ultrapure water; continuously passing a water sample through the small column to keep the overflowing speed of the water sample below 5mL/min, and cleaning the small column by using 10mL of ultrapure water after the sample loading of the sewage water sample is finished; continuously performing vacuum pumping for 20min by using a solid phase extraction device to fully dry the small column, and then putting the small column into a centrifugal machine for further dehydration and drying; the column was eluted with 20mL of methanol and the eluate was collected, followed by the same procedure as the invention.
The TOC value was measured using an Shimadzu TOC-VCPN instrument; scanning spectra were measured using Shimadzu UV-1800; measuring the three-dimensional fluorescence by using a Hitachi F-7000 instrument; measuring the trace organic pollutants by using a UPLC-Xevo TQ MS instrument; acute toxicity test of luminescent bacteria (Virbio fischeri) the test was carried out using the microplate toxicity method, according to the modified ISO11348 method.
Organic matters obtained by the method and the common methanol method are contrastively analyzed, and five parameters including TOC, UV-vis, 3DEEM, trace organic pollutants, acute toxicity of luminous bacteria and the like are included, as shown in figures 2-5. The total TOC recovery rate of the method is 89%, and can be improved by 50% compared with the TOC recovery rate of a methanol method; the organic matter obtained by the method achieves the desalting effect, and simultaneously, the ultraviolet-visible light region spectral parameters of the organic matter and the organic matter in the raw water can be highly matched; three-dimensional fluorescence spectrum parameters of the organic matter obtained by the method can show that compared with a methanol method, the recovery rate of the obtained organic matter fluorescence substances is greatly improved; the quantitative detection of trace organic pollutants by the organic matters obtained by the method shows that compared with a methanol method, the detection concentration of a single substance is obviously improved, and the total detection concentration is improved by 86 percent; compared with a methanol method, when the inhibition rate is 50%, the concentration multiple of the methanol method is 55.5 times, but the concentration multiple required by the method is only 15.5 times, so that the volume of a sewage water sample in biotoxicity detection is greatly reduced.
Claims (8)
1. The organic matter enrichment and concentration method based on low-pollution water body chemistry and biological analysis is characterized by comprising the following steps of:
step one, pretreating a sewage sample:
step two, enriching and concentrating organic matters:
utilizing a solid phase extraction device to enrich and concentrate organic matters in a water sample:
(1) connecting a C18 solid phase extraction column and an HLB solid phase extraction column in series;
(2) sequentially carrying out activation cleaning on 10mL of mixed solution of dichloromethane and normal hexane with a volume ratio of 1:1, 10mL of methanol and 10mL of ultrapure water, wherein the pH value of the ultrapure water is acidified to 2.5, and sequentially passing through a C18 and HLB solid-phase extraction small column;
(3) sequentially passing the sewage sample obtained in the step one through a solid phase extraction column formed by serially connecting C18 and HLB, and adjusting the pressure of a vacuum pump;
(4) after the sample loading of the sewage water sample is finished, using 10mL of ultrapure water, and cleaning the tandem small column by using the ultrapure water after acidification with the pH value of 2.5;
(5) continuously performing vacuum pumping by using a solid phase extraction device to fully dry the small columns connected in series, and then putting the small columns into a centrifugal machine for further dehydration and drying;
step three, preparing a concentrated organic solution:
(1) sequentially passing 10mL of methanol, 10mL of dichloromethane and n-hexane mixed liquor with the volume ratio of 1:1 through the dried solid phase extraction small column respectively, and collecting eluent;
(2) blowing the eluent to about 2mL by using high-purity nitrogen under the water bath condition, then concentrating all the eluent into a glass test tube, and blowing the eluent to a fully dry state by using high-purity nitrogen;
(3) dissolving the obtained solid organic matter by using phosphate solution, wherein the final concentration multiple of the organic matter is determined according to the biological toxicity detection result;
(4) and dissolving the obtained solid organic matters by using an organic solvent, and using the dissolved solid organic matters for subsequent sample loading detection of a liquid chromatography-mass spectrometer or a gas chromatography-mass spectrometer.
2. The method for enriching and concentrating organic matters based on the chemical and biological analysis of the low-pollution water body as claimed in claim 1, wherein in the step one; the water sample pretreatment specifically comprises the combined use of sewage water sample acidification and the addition of a 5% methanol organic solvent.
3. According toThe method for enriching and concentrating organic matters based on the chemical and biological analysis of low-pollution water bodies as claimed in claim 1, wherein in the first step, the glass fiber filter membrane is 0.7/0.45 μm, and H is2SO4The concentration was 2M.
4. The method for enriching and concentrating the organic matters based on the chemical and biological analysis of the low-pollution water body as claimed in claim 1, wherein in the second step, the volume ratio of the mixed solution of the dichloromethane and the n-hexane is 1:1, the methanol is used in a ratio of the dichloromethane to the n-hexane, the volume ratio of the mixed solution of the dichloromethane and the n-hexane is 1:1, and the polarity coverage range of the activated solvent is 0.06 of the n-hexane → 3.4 of the dichloromethane → 6.6 of the methanol.
5. The method for enriching and concentrating organic matters based on the chemical and biological analysis of the low-pollution water body as claimed in claim 1, wherein the water sample overflowing speed in the second step is kept below 5mL/min, and the vacuum pumping is continued for 20min by using a solid phase extraction device.
6. The method for enriching and concentrating the organic matters based on the chemical and biological analysis of the low-pollution water body as claimed in claim 1, wherein in the third step, the volume ratio of the mixed solution of dichloromethane and n-hexane is 1:1, and the volume ratio of the mixed solution of methanol and n-hexane is 1: 1.
7. The method for enriching and concentrating organic matters based on the chemical and biological analysis of the low-pollution water body as claimed in claim 1, wherein the water bath condition in the third step is 40 ℃, the phosphate solution is 5mM, and the pH value is 8.
8. The organic matter enrichment and concentration method based on the chemical and biological analysis of the low-pollution water body according to claim 1, characterized in that the first step is to collect a sewage water sample according to a standard sampling method and to treat the water sample as follows:
(1) filtering a certain amount of sewage water sample by a glass fiber filter membrane by using a large-volume filtering device;
(2) the filtered water sample is stored in a glass containerIn the vessel, use H2SO4Acidifying the pH of the water sample to 2.5;
(3) adding a certain amount of chromatographic grade methanol solution into a water sample to ensure that the volume fraction of the methanol solution is 5 percent.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112730650A (en) * | 2020-12-15 | 2021-04-30 | 湖南微谱检测技术有限公司 | Method for enriching ultra-trace organic matters in large-volume water |
| CN113092705A (en) * | 2021-04-09 | 2021-07-09 | 西安建筑科技大学 | Rapid, simple and convenient pretreatment method for biotoxicity detection of low-toxicity sample |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112730650A (en) * | 2020-12-15 | 2021-04-30 | 湖南微谱检测技术有限公司 | Method for enriching ultra-trace organic matters in large-volume water |
| CN113092705A (en) * | 2021-04-09 | 2021-07-09 | 西安建筑科技大学 | Rapid, simple and convenient pretreatment method for biotoxicity detection of low-toxicity sample |
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Application publication date: 20200519 |