CN108129547A - A kind of method for extracting zoogloea extracellular polymeric - Google Patents
A kind of method for extracting zoogloea extracellular polymeric Download PDFInfo
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Abstract
The present invention provides a kind of methods of high efficiency extraction extracellular polymeric substances from activated sludge, and including cleaning, centrifugal process extraction slime layer, supercentrifugal process extracts loose associativity extracellular polymeric (LB EPS), formaldehyde-acetic acid-potassium acetate heating extracts type extracellular polymeric (TB EPS) of combining closely.The beneficial effects of the invention are as follows：Method of the present invention is capable of easy, quickly extraction activated sludge extracellular polymeric, and when can avoid dissolution of cellular content to extracting solution and extraction the decomposition, to greatest extent guarantee extract of caused protein and polysaccharide purity.
Description
Technical field
The present invention relates to a kind of extracting methods of activated sludge zoogloea extracellular polymeric, and it is extracellular to can be applied to activated sludge
The extraction of polymer and the research of character.In terms of being particularly applicable to sewage disposal, belong to technical field of bioengineering.
Background technology
Extracellular polymeric (Extracellular Polymeric Substances, EPS) is that microbial cell is being metabolized
The general name of extracellular polymer substance secreted in the process, mainly including polysaccharide, nucleic acid and lipid etc..Extracellular polymeric is activity
The important component of sludge, it has certain hydrophobicity, can be combined with cation, to sludge surface charge, sedimentation, flocculation and takes off
It is aqueous that all there is important influence.Study influence and its quantitative analysis of the extracellular polymeric to Sludge Property, it is necessary first to one
It is suitble to the method for extraction sludge extracellular polymeric.Extracellular polymeric extraction method have very much, mainly include physics extraction method with
Chemical leaching test.It in extraction process, should ensure to extract the total amount of extracellular polymeric, ensure that biological cell is not broken again
It is bad, therefore the extraction of sludge extracellular polymeric is influenced by several factors.
The extraction of EPS is that the important prerequisite of ingredient and functional analysis is carried out to it, and standard method is not yet established.Same Way
To different sample EPS extraction efficiencies difference, distinct methods extraction same sample EPS efficiency also differs greatly.There is no a kind of sides
Method can all extract sample EPS, and all methods can all cause cell rupture to a certain extent.And evaluate an extracting method
It is whether suitable, not only to consider EPS yield height, also to consider destruction of the extracting method to macromolecular substances in cell and EPS
Degree and whether interfere the correlative studys such as subsequent measurements.
Invention content：
The defects of it is an object of the invention to overcome the prior art, provides one kind and does not destroy cell microstructure, chemistry examination
Smaller, the high Undec extracellular polymeric substances from activated sludge extracting method of extract purity is interfered in agent.
The present invention provides a kind of extracellular polymeric extracting solution, is by weight：5-6 parts of titanium dioxide, 3.6% formaldehyde are molten
5-6 parts of liquid, acetic acid -10-15 parts of potassium acetate solution that pH is 5.5,20-25 parts of quartz sand.
The application method of the extracellular polymeric extracting solution, the additive amount of every liter of sludge are：40-52g/L sludge.
The present invention provides a kind of method of high efficiency extraction zoogloea extracellular polymeric, comprises the steps of：
Cleaning
It is in situ to obtain mud sample, its volume is measured, is covered on Buchner funnel after filter paper is soaked, sludge is placed on
On filter paper, suction filtration machine is opened, sodium chloride solution is added in funnel, sludge is cleaned, until the solution to leak down becomes clarification；
Centrifugal process extracts slime layer
Treated, and sludge adds in sodium chloride solution recovery original volume, is distributed into centrifuge tube, in the centrifuge of the slow-speed of revolution
Interior centrifugation, extraction supernatant are slime layer.
Supercentrifugal process extracts loose associativity extracellular polymeric (LB-EPS)
Centrifuge tube sludge is restored to original volume using sodium chloride solution.Using 120W ultrasounds by Treatment of Sludge 1.5-
2min.Using being centrifuged under conditions of low temperature, high speed in supercentrifuge, extraction supernatant is loose mating type extracellular polymeric
(LB-EPS)；
Formaldehyde-acetic acid-potassium acetate heating extracts type extracellular polymeric (TB-EPS) of combining closely
The sludge obtained in step (3) is all added in beaker, sodium chloride solution is added in and restores to original volume；Addition carries
Liquid is taken, sludge is put into concussion and cultivate case with 25-27 DEG C, 150-200r/min shake culture 15-20min, is taken out.It is described to carry
The liquid is taken to be：Titanium dioxide 5-6g/L sludge, 3.6% formalin 5-6mL/L sludge, acetic acid-potassium acetate solution that pH is 5.5
10-15mL/L sludge, quartz sand 20-25g/L sludge.
It by sludge heating water bath 20min in beaker, is sub-packed in centrifuge tube after cooling, low temperature is high under conditions of 1-4 DEG C
Speed centrifugation 20-22min.It is (TB-EPS) to extract supernatant.
The slime layer of extraction, LB-EPS are merged into mixing with TB-EPS, using membrane filtration, as extracellular polymeric extracts
Liquid.Extracting solution should be stored refrigerated under the conditions of 1-4 DEG C.
Wherein, it is preferred that in step (4), the preparation method for acetic acid-potassium acetate solution that pH is 5.5 is：By 41g acetic acid
Potassium is settled to 500mL respectively with 29.5mL glacial acetic acids, by potassium acetate solution and acetic acid solution according to 9:1 volume ratio mixing, it is close
Envelope preserves, as the potassium acetate solution of pH=5.5.
Preferably, the preferred graininess anaerobic ammonium oxidation sludge of step (1) described sludge or graininess anaerobic sludge or biology
The above-mentioned sludge of film form.
Preferably, the sodium chloride solution preferred concentration described in step (1) is the sodium chloride solution of 0.50%-0.80%.It takes out
The pressure of filter is preferably 0.03-0.04MPa.
Preferably, the centrifugal speed of step (2) described centrifuge is preferably 550-650r/min, centrifugation time 10-
15min, centrifuging temperature are preferably 28 DEG C.
Preferably, the sodium chloride that step (3), the sodium chloride solution preferred concentration described in (4) are 0.05%-0.10% is molten
Liquid.
Preferably, the temperature of the centrifuge in step (3) is preferably 3-5 DEG C, and centrifugal speed is preferably 5000-6000r/
Min, centrifugation time are preferably 8-10min.
Preferably, the preferred 50-100 mesh of quartz sand size in step (4), additive amount are 20-25g/L sludge
Preferably, the water bath heating temperature in step (5) is preferably 60-70 DEG C.
Preferably, the centrifuge speed in step (5) is preferably 8000-9000r/min, and centrifugation duration is preferably 18-
20min。
Preferably, filtering uses 0.40-0.45 μm of water system filter membrane pressure filtration in step (6).
The present invention concrete principle be：
The present invention obtains caducous zoogloea slime layer using low-speed centrifugal, and refrigeration medium-speed centrifuge extraction is loose to be combined
Type extracellular polymeric；By controlling temperature, control centrifugal speed, the structure of cell interior is protected to greatest extent.It is innovative
Formalin and weak acetic acid-potassium acetate solution help of combination low concentration dissociate the type born of the same parents that combine closely on zoogloea surface
Outer polymer, and use quartz sand with titanium dioxide as rubbing agent, it is by shearing, rubbing action that zoogloea surface is close
The extracellular polymeric of attachment strips down.The larger quartz sand of particle and the smaller titania powder collective effect of particle, make
Extracellular polymeric detaches more thorough.
Compared with prior art, the present invention has the following advantages：
The physical method of extraction each section extracellular polymeric announced of the present invention, by cleaning, centrifuging, dissociating etc. just
Method realizes the extraction respectively to activated sludge each section extracellular polymeric, convenient for each group of separated research extracellular polymeric
Into part.Efficiently separating, and do not injure cell itself for extracellular polymeric and bacterium ontology is realized, is avoided intracellular various
The substances such as polysaccharide, protein, DNA spilling interferes.Also avoiding a large amount of NaOH and methanol leads to albumen in extracting solution simultaneously
The inactivation of the bioactive substances such as matter convenient for subsequent research and measures.
Description of the drawings：
Fig. 1-Fig. 4 is the amount of the extracellular polymeric of various methods extraction and its constituent content comparison.
Fig. 1 is the comparison of three kinds of EPS total amounts of extraction.
Fig. 2 is the comparison of polyoses content in the three kinds of EPS extracted.
Fig. 3 is the comparison of protein content in the three kinds of EPS extracted.
Fig. 4 is the comparison of DNA content in the three kinds of EPS extracted.
Specific embodiment
Following embodiment is the further explanation to the present invention, but the present invention is not limited thereto.
Embodiment 1
The extracellular polymeric of certain black anaerobic grain sludge is extracted using this patent the method and measures its ingredient.Specifically
Operating procedure is as follows：
Mud sample is obtained out of reactor, measures its volume as 30mL.It is covered on Buchner funnel after filter paper is soaked,
Sludge is placed on filter paper, opens suction filtration machine, it is 0.03MPa to adjust suction filtration machine pressure.0.8% is slowly poured into funnel
Sodium chloride solution, sludge is cleaned.
Treated, and sludge takes out, and is fitted into beaker, adds in sodium chloride solution and restores volume to 30mL.It is distributed into
In 1.5mL centrifuge tubes, centrifuge is put into, adjustment centrifugal rotational speed is 600r/min, setting time 15min, and extraction supernatant is
Slime layer.
Sludge in centrifuge tube is taken out, is restored sludge volume to 30mL using sodium chloride solution.At 120W ultrasounds
Manage sludge 2min.Centrifuge speed is set as 5500r/min, centrifuging temperature is set as 4 DEG C, and centrifugation time is set as 10min,
Extraction supernatant is loose mating type extracellular polymeric (LB-EPS).
The sludge obtained in step (3) is all added in beaker, sodium chloride solution is added in and restores to 30mL.Add in dioxy
Change the acetic acid-potassium acetate solution 0.3mL sludge and 75 mesh quartz sand 0.75g that titanium 0.15g, 3.6% formaldehyde 0.15mL, pH are 5.5.
Sludge is put into concussion and cultivate case with 27 DEG C, 150r/min shake culture 15min, is taken out.
Beaker is put into 60 DEG C of heating water bath 20min, is sub-packed in after cooling in 1.5mL centrifuge tubes, under conditions of 4 DEG C with
9000r/min centrifuges 20min.It is (TB-EPS) to extract supernatant.
The slime layer of extraction, LB-EPS are uniformly mixed with TB-EPS, using syringe and 0.45 μm of membrane filtration, as
The EPS extracting solutions of the anaerobic grain sludge.This extracting solution is positioned over stored refrigerated under 4 DEG C of environment, measures its protein, polysaccharide
With the content of DNA.The wherein measure of protein measures its content using bovine serum albumin as standard specimen, with Coomassie Brilliant Blue；Polysaccharide
Measure using glucose as standard specimen, measured with anthrone colorimetry；The content of DNA using standard DNA solution as standard specimen, with diphenylamines-
Acetaldehyde spectrophotometry measures its content.(note：Measure DNA before need to be purified with phenol extraction method, remove sample solution in protein and
Polysaccharide.)
Embodiment 2
The extracellular polymeric of certain red anaerobic ammonium oxidation granular sludge is extracted using this patent the method and measure its into
Point.Concrete operation step is as follows：
Mud sample is obtained out of anaerobic ammonia oxidation reactor, measures its volume as 30mL.It is covered in after filter paper is soaked
On Buchner funnel, sludge is placed on filter paper, opens suction filtration machine, it is 0.03MPa to adjust suction filtration machine pressure.Slowly to funnel
0.8% sodium chloride solution is inside poured into, sludge is cleaned.
Treated, and sludge takes out, and is fitted into beaker, adds in sodium chloride solution and restores volume to 30mL.It is distributed into
In 1.5mL centrifuge tubes, centrifuge is put into, adjustment centrifugal rotational speed is 600r/min, setting time 15min, and extraction supernatant is
Slime layer.
Sludge in centrifuge tube is taken out, is restored sludge volume to 30mL using sodium chloride solution.At 120W ultrasounds
Manage sludge 2min.Centrifuge speed is set as 5500r/min, centrifuging temperature is set as 4 DEG C, and centrifugation time is set as 10min,
Extraction supernatant is loose mating type extracellular polymeric (LB-EPS).
The sludge obtained in step (3) is all added in beaker, sodium chloride solution is added in and restores to 30mL.Add in dioxy
Change the acetic acid-potassium acetate solution 0.3mL sludge and 75 mesh quartz sand 0.9g that titanium 0.16g, 3.6% formaldehyde 0.15mL, pH are 5.5.
Sludge is put into concussion and cultivate case with 27 DEG C, 180r/min shake culture 15min, is taken out.
Beaker is put into 60 DEG C of heating water bath 20min, is sub-packed in after cooling in 1.5mL centrifuge tubes, under conditions of 4 DEG C with
9000r/min centrifuges 20min.It is (TB-EPS) to extract supernatant.
The slime layer of extraction, LB-EPS are uniformly mixed with TB-EPS, using syringe and 0.45 μm of water system membrane filtration,
The as EPS extracting solutions of the anaerobic grain sludge.This extracting solution is positioned over it is stored refrigerated under 4 DEG C of environment, measure its protein,
The content of polysaccharide and DNA.The wherein measure of protein measures its content using bovine serum albumin as standard specimen, with Coomassie Brilliant Blue；
The measure of polysaccharide is measured using glucose as standard specimen with anthrone colorimetry；The content of DNA is using standard DNA solution as standard specimen, with hexichol
Amine-acetaldehyde spectrophotometry measures its content.(note：It need to be purified before measuring DNA with phenol extraction method, remove albumen in sample solution
Matter and polysaccharide.)
Comparative example 1
Using traditional centrifugation-ultrasonic method the red anaerobic grain dirt identical with embodiment two is extracted with formaldehyde-NaOH methods
The extracellular polymeric of mud simultaneously measures its ingredient.Concrete operation step is as follows：
Mud sample after cleaning under the conditions of 600r/min is centrifuged into 15min, takes its supernatant through 0.45 μm of filter membrane mistake
Filter to obtain slime layer.
By remaining sludge 0.85%NaCl solution suspensions to original volume, 2min is ultrasonically treated with 120W, then
The shake culture 10min under the conditions of 28 DEG C, 180r/min, with 5000r/min, 4 DEG C of centrifugation 20min, collection supernatant is LB-
EPS。
Excess sludge is poured into beaker, original volume is suspended to 0.85% sodium chloride, is ultrasonically treated with 120W
2min, the shake culture 10min under the conditions of 28 DEG C, 180r/min.
By (3) step, treated that sludge is all collected into beaker, and the formaldehyde of 0.06mL36.5% is added in into beaker,
1h is vibrated in 28 DEG C, 180r/min shaking tables, 1mol/LNaOH solution is added and adjusts pH to 11, vibrate 3h in shaking table, then
20min is centrifuged under conditions of 20000r/min, 4 DEG C, collection supernatant is TB-EPS.
Measure the content of the extracting solution protein, polysaccharide and DNA.Wherein the measure of protein is using bovine serum albumin as mark
Sample measures its content with Coomassie Brilliant Blue；The measure of polysaccharide is measured using glucose as standard specimen with anthrone colorimetry；DNA's contains
Amount measures its content using standard DNA solution as standard specimen with diphenylamines-acetaldehyde spectrophotometry.(note：It need to be taken out before measuring DNA with phenol
Formulation purifies, and removes protein and polysaccharide in sample solution.)
Comparison more than case, in the extracellular polymeric extracted using method provided by the invention contained DNA content compared with
It is few, respectively 2.32mggss-1 and 2.51mggss-1, and use the born of the same parents of the anaerobic ammonium oxidation sludge of traditional method for extracting
Outer polymer DNA is up to 7.9mmggss-1, illustrates conventional method to the destructions of cellular entities when this patent the method
By force.The extracellular extract total amount of anaerobic ammonium oxidation sludge that this patent the method obtains is 33.65mggss-1, although than passing
The extracellular polymeric that system formaldehyde-NaOH methods are extracted is few (45.31mggss-1), but its damage to thalline is smaller, comprising
Cell interior substance is less.Since anaerobic ammonia oxidizing bacteria is cultivated using artificial wastewater, in living environment, organic matter is very
It is few, so the polysaccharide and protein contained by its extracellular polymeric are all than relatively low.And commonly the anaerobic sludge speed of growth is fast, has in height
It survives in machine matter environment, so the content of protein and polysaccharide is higher, is higher by 35.23% and 26.01% respectively.
Since common anaerobic sludge is largely heterotroph bacterium, breeding is fast, and for handling high concentrated organic wastewater, so
Its extracellular polymeric aggregate velocity is higher, and the content of organic matter is high；Anaerobic ammonia oxidizing bacteria is autotrophic type bacterium, and breeding is slow, and for locating
Inorganic wastewater is managed, so its extracellular polymeric aggregate velocity is relatively low, the content of organic matter is low.Although use detesting for traditional method for extracting
Anaerobic ammonium oxidation extracellular polymeric substances from activated sludge extracted amount is larger, but in its extract DNA content it is higher, illustrate its eucaryotic cell structure
It is destroyed, the content outflow in cell polluted the extracting solution of extracellular polymeric, cause measurement result inaccurate.Make
It is compared with the extracellular polymeric that is extracted of novel method that this patent provides and conventional method, although the content of each component and total
Measure slightly lower, but wherein the content of DNA only has 1/3rd of traditional method for extracting liquid to a quarter, illustrates described in this patent
Method can be effectively protected eucaryotic cell structure while extracellular polymeric is extracted, ensure extraction extracellular polymeric it is pure
Just, the influence of experiment is preferably minimized.
Claims (10)
1. a kind of extracellular polymeric extracting solution, is by weight：5-6 parts of titanium dioxide, 3.6% 5-6 parts of formalin, pH are
5.5 acetic acid -10-15 parts of potassium acetate solution, 20-25 parts of quartz sand.
2. the application method of extracellular polymeric extracting solution as described in claim 1, the additive amount of every liter of sludge are：40-52g/L
Sludge.
3. a kind of method of high efficiency extraction zoogloea extracellular polymeric, comprises the steps of：
Cleaning
It is in situ to obtain mud sample, its volume is measured, is covered on Buchner funnel after filter paper is soaked, sludge is placed on filter paper
On, suction filtration machine is opened, sodium chloride solution is added in funnel, sludge is cleaned, until the solution to leak down becomes clarification；
Centrifugal process extracts slime layer
Treated sludge adds in sodium chloride solution and restores original volume, is distributed into centrifuge tube, in the centrifuge of the slow-speed of revolution from
The heart, extraction supernatant are slime layer；
Supercentrifugal process extracts loose associativity extracellular polymeric (LB-EPS)
Centrifuge tube sludge is restored to original volume using sodium chloride solution.Using 120W ultrasounds by Treatment of Sludge 1.5-2min；Make
With being centrifuged under conditions of low temperature, high speed in supercentrifuge, extraction supernatant is loose mating type extracellular polymeric (LB-
EPS)；
Formaldehyde-acetic acid-potassium acetate heating extracts type extracellular polymeric (TB-EPS) of combining closely
The sludge obtained in step (3) is all added in beaker, sodium chloride solution is added in and restores to original volume；Add in extraction
Sludge is put into concussion and cultivate case with 25-27 DEG C, 150-200r/min shake culture 15-20min, takes out by liquid；The extraction
Liquid is：Titanium dioxide 5-6g/L sludge, 3.6% formalin 5-6mL/L sludge, acetic acid-potassium acetate solution that pH is 5.5
10-15mL/L sludge, quartz sand 20-25g/L sludge；
By sludge heating water bath 20min in beaker, be sub-packed in centrifuge tube after cooling, under conditions of 1-4 DEG C low-temperature and high-speed from
Heart 20-22min.It is (TB-EPS) to extract supernatant；
The slime layer of extraction, LB-EPS are merged into mixing with TB-EPS, use membrane filtration, as extracellular polymeric extracting solution.
Extracting solution should be stored refrigerated under the conditions of 1-4 DEG C.
4. the method for high efficiency extraction zoogloea extracellular polymeric as claimed in claim 3, which is characterized in that in step (4), pH
Preparation method for 5.5 acetic acid-potassium acetate solution is：41g potassium acetates and 29.5mL glacial acetic acids are settled to 500mL respectively,
By potassium acetate solution and acetic acid solution according to 9:1 volume ratio mixing, is sealed, as the potassium acetate solution of pH=5.5.
5. the method for high efficiency extraction zoogloea extracellular polymeric as claimed in claim 3, which is characterized in that step (1) is described
Concentration of sodium chloride solution be 0.50%-0.80% sodium chloride solution；The pressure of suction filtration machine is 0.03-0.04MPa；Step
(3), the concentration of sodium chloride solution described in (4) is the sodium chloride solution of 0.05%-0.10%.
6. the method for high efficiency extraction zoogloea extracellular polymeric as claimed in claim 3, which is characterized in that step (2) is described
The centrifugal speed of centrifuge is 550-650r/min, and centrifugation time 10-15min, centrifuging temperature is 28 DEG C.
7. the method for high efficiency extraction zoogloea extracellular polymeric as claimed in claim 3, which is characterized in that in step (3)
The temperature of centrifuge is 3-5 DEG C, centrifugal speed 5000-6000r/min, centrifugation time 8-10min.
8. the method for high efficiency extraction zoogloea extracellular polymeric as claimed in claim 3, which is characterized in that in step (4)
Quartz sand size 50-100 mesh, additive amount are 20-25g/L sludge.
9. the method for high efficiency extraction zoogloea extracellular polymeric as claimed in claim 3, which is characterized in that in step (5)
Water bath heating temperature is 60-70 DEG C.Centrifuge speed in step (5) is 8000-9000r/min, a length of 18- during centrifugation
20min。
10. the method for high efficiency extraction zoogloea extracellular polymeric as claimed in claim 3, which is characterized in that mistake in step (6)
Filter uses 0.40-0.45 μm of water system filter membrane pressure filtration.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109111499A (en) * | 2018-08-23 | 2019-01-01 | 暨南大学 | A method of extracting ground bacillus Extracellular polymers |
| CN109211738A (en) * | 2018-07-18 | 2019-01-15 | 广西大学 | The detection judgment method of anaerobic grain sludge calcification and active height in high calcium waste water |
| CN110451762A (en) * | 2019-07-05 | 2019-11-15 | 重庆大学 | A kind of extracting method of extracellular polymeric substances from activated sludge |
| CN112898374A (en) * | 2021-02-05 | 2021-06-04 | 中南大学 | Method for extracting extracellular polymer of bacterial-algae symbiotic system |
| CN114797777A (en) * | 2022-04-28 | 2022-07-29 | 南京大学 | Preparation method of sludge-based biochar loaded nano-iron based on extracellular polymer regulation |
| CN114890636A (en) * | 2022-05-19 | 2022-08-12 | 浙江工业大学 | Activated sludge extracellular polymer extraction method based on divalent cation complexation |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010142004A8 (en) * | 2009-06-10 | 2011-11-24 | Katholieke Universiteit Leuven | Controlled biosecure aquatic farming system in a confined environment |
| CN104316502A (en) * | 2014-10-18 | 2015-01-28 | 北京工业大学 | Method for optimizing ultrasonic conditions and extracting extracellular polymeric substances through three-dimensional fluorescence spectrum |
| WO2015154731A1 (en) * | 2014-04-09 | 2015-10-15 | Vakos Xt A.S. | A formulation in the form of a suspension for the treatment of blood-sucking parasite |
| CN107011410A (en) * | 2017-03-01 | 2017-08-04 | 中山大学 | A kind of use quartz sand extracts the method and its application of sludge extracellular polymeric |
-
2017
- 2017-11-21 CN CN201711167014.2A patent/CN108129547B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2010142004A8 (en) * | 2009-06-10 | 2011-11-24 | Katholieke Universiteit Leuven | Controlled biosecure aquatic farming system in a confined environment |
| WO2015154731A1 (en) * | 2014-04-09 | 2015-10-15 | Vakos Xt A.S. | A formulation in the form of a suspension for the treatment of blood-sucking parasite |
| CN104316502A (en) * | 2014-10-18 | 2015-01-28 | 北京工业大学 | Method for optimizing ultrasonic conditions and extracting extracellular polymeric substances through three-dimensional fluorescence spectrum |
| CN107011410A (en) * | 2017-03-01 | 2017-08-04 | 中山大学 | A kind of use quartz sand extracts the method and its application of sludge extracellular polymeric |
Non-Patent Citations (2)
| Title |
|---|
| GUANG-HUI YU等: "Extracellular proteins, polysaccharides and enzymes impact on sludge aerobic digestion after ultrasonic pretreatment", 《WATER RESEARCH》 * |
| 曹秀芹 等: "胞外聚合物（EPS）构成的影响因素分析", 《环境科学与技术》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109211738A (en) * | 2018-07-18 | 2019-01-15 | 广西大学 | The detection judgment method of anaerobic grain sludge calcification and active height in high calcium waste water |
| CN109211738B (en) * | 2018-07-18 | 2021-04-13 | 广西大学 | Method for detecting and judging calcification and activity of anaerobic granular sludge in high-calcium wastewater |
| CN109111499A (en) * | 2018-08-23 | 2019-01-01 | 暨南大学 | A method of extracting ground bacillus Extracellular polymers |
| CN110451762A (en) * | 2019-07-05 | 2019-11-15 | 重庆大学 | A kind of extracting method of extracellular polymeric substances from activated sludge |
| CN110451762B (en) * | 2019-07-05 | 2020-09-18 | 重庆大学 | Extraction method of activated sludge extracellular polymers |
| CN112898374A (en) * | 2021-02-05 | 2021-06-04 | 中南大学 | Method for extracting extracellular polymer of bacterial-algae symbiotic system |
| CN112898374B (en) * | 2021-02-05 | 2023-02-03 | 中南大学 | Method for extracting extracellular polymer of bacterial-algae symbiotic system |
| CN114797777A (en) * | 2022-04-28 | 2022-07-29 | 南京大学 | Preparation method of sludge-based biochar loaded nano-iron based on extracellular polymer regulation |
| CN114890636A (en) * | 2022-05-19 | 2022-08-12 | 浙江工业大学 | Activated sludge extracellular polymer extraction method based on divalent cation complexation |
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