CN108042968A - A kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation - Google Patents
A kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation Download PDFInfo
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Abstract
The invention discloses a kind of preparation method of the reagent for polycyclic aromatic hydrocarbon selective degradation, including：The synthesis of lumichrome, once purifying, secondarily purified, chlorella protein liquid the extraction of lumichrome, the purifying of protein liquid, the compounding of reagent of lumichrome, the reagent is using photopigment lumichrome as probe, compound as primer using chlorella liquid of protease, addition stabilizer is prepared.It has the beneficial effect that：Using photopigment lumichrome as polycyclic aromatic hydrocarbon detection probe, polycyclic aromatic hydrocarbon can efficiently be positioned, improve the efficiency for polycyclic aromatic hydrocarbon selective degradation, the chlorella protein liquid of reagent release simultaneously can be effectively broken for the benzene ring structure of polycyclic aromatic hydrocarbon, it is the substances such as aldehyde, alcohol by degrading polycyclic aromatic hydrocarbons, is eliminated so as to targetedly complete degradation to it.
Description
Technical field
The present invention relates to Environmental Engineering Material technical field, more particularly, to a kind of for polycyclic aromatic hydrocarbon selective degradation
The preparation method of reagent.
Technical background
At present, the enrichment of polycyclic aromatic hydrocarbon can be directly resulted in once Oil spills occurs in marine environment, soil environment, into
And threat is generated to life entity.The hormonal action of polycyclic aromatic hydrocarbon can carcinogenic, teratogenesis, mutagenesis, generate genotoxicity, to human body
Immune system damages.Therefore, how pointedly, efficiently polycyclic aromatic hydrocarbon contained in degradation of oils, barrier human body are taken the photograph
Entering the channel of polycyclic aromatic hydrocarbon is then particularly important.
Mainly there is Physical currently for the processing method of polycyclic aromatic hydrocarbon（Heating, coagulant sedimentation, absorption method）, chemistry
Method（Photooxidation method, chemical agent oxidizing process）, microbial method.In view of physics, chemical method handle polycyclic aromatic hydrocarbon, removal is not thorough
While the hidden danger of chemical residual is also resulted in environment, microbial method is put forth effort in research at present.The microbiological treatment delivered is more
The related data of cycloaromatics shows there is complicated structure due to polycyclic aromatic hydrocarbon, therefore its degradation rate is not generally high, and related micro-
Biology also has certain limitation when for the degradation of polycyclic aromatic hydrocarbon, i.e., in most cases, a kind of microorganism specific can only drop
Solve a kind of polycyclic aromatic hydrocarbon.
The content of the invention
It is an object of the invention to provide a kind of preparation method of the reagent for polycyclic aromatic hydrocarbon selective degradation, preparation sides
Method is simple and practicable, safety and environmental protection, and reagent has polycyclic aromatic hydrocarbon guidance quality and special degradability, while has both less toxic, environmentally friendly, steady
Calmly, the advantages of action condition is mild, contaminant degradation is thorough, and to environment non-secondary pollution.
The problem of present invention in background technology for mentioning, the technical solution taken is：One kind is special for polycyclic aromatic hydrocarbon
Property degradation reagent preparation method, the once purifying of synthesis, lumichrome including lumichrome, dimethyl are different
Secondarily purified, chlorella protein liquid the extraction of alloxazine, the purifying of protein liquid, the compounding of reagent, specifically include following steps：
The synthesis of lumichrome：6, the 7 dimethyl -9- formic acid ylmethyl isoalloxazines that 1.0-1.2g is dried（FMF）Solid
It is placed in beaker, the boric acid solution of addition 30-32mL 0.10-0.12M and the sodium hydroxide of 130-135mL 0.1-0.12M are molten
Liquid is protected from light and is stirred to react 90-100 minutes to obtain lumichrome, reaction principle such as Fig. 1；It is being protected from light, under alkaline condition, by FMF
It sloughs aldehyde-base and lumichrome is prepared, preparation method is more mild, and reaction process is easily-controllable, simple for process；
The once purifying of lumichrome：Filtrate is placed in separatory funnel by filtering reacting liquid, adds in dichloromethane extraction,
Extraction is until extract liquor is colourless repeatedly；Extract liquor rotary evaporation is concentrated with Rotary Evaporators at a temperature of 38-40 DEG C,
Concentrate final volume is 18-20mL；Selecting dichloromethane, toxicity is relatively low, and effect of extracting is preferable, is into one as extractant
The purifying of step is prepared；
Lumichrome it is secondarily purified：Thin-layer chromatography separation is done to concentrate, thickness of thin layer 0.18-0.25mm passes through
Gel imager observes fluorescence, then uses column chromatography concentrate, and the stationary phase of column chromatography is silica gel, and mobile phase is that weight ratio is
6-8:3-5:1 n-butanol, methanol, distilled water obtain the solution of lumichrome after chromatography；By solution in 70-74 DEG C of rotation
Turn evaporation, obtain concentrate, then be placed in vacuum drying chamber and be dried to obtain light yellow lumichrome solid with 55-58 DEG C；
Thin-layer chromatography is suitable for that micro, volatility is smaller, the substance separation of heatproof, according to lumichrome and impurity in stationary phase
Progress column chromatography for separation different with distribution coefficient in mobile phase, the content of lumichrome obtains after isolating and purifying twice
It greatly improves；
The extraction of chlorella protein liquid：By chlorella centrifugal concentrating 8-10 minutes under the conditions of 2-4 DEG C, 6000-8000r/min,
Concentrate is subjected to breaking-wall cell in Ultrasonic Cell Disruptor, cell crushing instrument power is 1.0-1.2kW, and probe amplitude is 35-
38%, it is 30-45 minute to crush the time, it is broken after obtained by liquid be placed in 2-4 DEG C, centrifuge 15- under the conditions of 10000-13000r/min
20 minutes, it was crude protein liquid to take supernatant；Ultrasonic Cell Disruptor is selected to crush chlorella, shell-broken effect is preferable, will not be to activity
Substance impacts, and will not add new polluter, is conducive to the extraction and purification of chlorella albumen；
The purifying of protein liquid：Protein liquid is placed in saturated ammonium sulfate solution, is chromatographed through DEAE Sepharose Fast Flow
Column, with sodium chloride buffer gradient elution, NaCl concentration gradient 100mmol/L, 200 mmol/L, 400 mmol/L, 700
mmol/L；Merge the eluent with enzyme activity, be the phosphoric acid of 7.4-7.6 by 1.0-1.3mM, pH of concentration at a temperature of 0-4 DEG C
Buffer solution dialysed overnight obtains protein liquid after purification；With sodium chloride buffer gradient elution, not only separation accuracy is high, efficiency
It is higher, and new pollutant will not be introduced；
The compounding of reagent：50-55mg lauryl sodium sulfate, 0.5-0.6g lumichrome powder are added in protein liquid,
It stirs evenly, stands 30-45 minutes, add in stabilizer to get for the reagent for more changing aromatic hydrocarbons selective degradation；With photopigment two
Methyl isoalloxazine is polycyclic aromatic hydrocarbon detection probe, can efficiently position polycyclic aromatic hydrocarbon, is improved for polycyclic aromatic hydrocarbon selective degradation
Efficiency, while the chlorella protein liquid of reagent release can be effectively broken for the benzene ring structure of polycyclic aromatic hydrocarbon, by polycyclic virtue
Alkane degradation is the substances such as aldehyde, alcohol, is eliminated so as to targetedly complete degradation to it.
Preferably, the stabilizer in the compounding step of reagent is 10-12% glycerine, 2.0-2.3mMEDTA, 0. 5-
The mixed liquor of 0.6mMDTT, pH 7.6-7.8.
Compared with prior art, the advantage of the invention is that：
1）Using photopigment lumichrome as polycyclic aromatic hydrocarbon detection probe, polycyclic aromatic hydrocarbon can be efficiently positioned, is improved for polycyclic
The efficiency of aromatic hydrocarbons selective degradation, at the same reagent release chlorella protein liquid can effectively for polycyclic aromatic hydrocarbon benzene ring structure into
Degrading polycyclic aromatic hydrocarbons are the substances such as aldehyde, alcohol by row fracture, are eliminated so as to targetedly complete degradation to it.
2）It is easily obtained for extracting the chlorella of protein liquid from environment, and training method is easy to operate, convenient for breeding, into
This is cheap, and in actual environment reparation operation, because of its less toxic characteristic and action condition is mild, is more conducive to actual repair effect
Prediction, and degrade different from general chemical agent, the present invention is more thorough to the degradation of pollutant based on chlorella protein liquid
Bottom.
Description of the drawings
Fig. 1 is the reaction principle schematic diagram for preparing lumichrome in the present invention by FMF.
Specific embodiment
The present invention program is described further below by drawings and examples：
Embodiment 1：
A kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation, comprises the following steps：
The synthesis of lumichrome：6, the 7 dimethyl -9- formic acid ylmethyl isoalloxazine solids that 1g is dried is taken to be placed in beaker,
The boric acid solution of 30mL 0.10M and the sodium hydroxide solution of 130mL 0.10M are added in, is protected from light and is stirred to react 90 minutes to obtain diformazan
Base isoalloxazine, reaction principle such as Fig. 1；It being protected from light, under alkaline condition, FMF is sloughed aldehyde-base is prepared lumichrome,
Preparation method is more mild, and reaction process is easily-controllable, simple for process；
The once purifying of lumichrome：Filtrate is placed in separatory funnel by filtering reacting liquid, adds in dichloromethane extraction,
Extraction is until extract liquor is colourless repeatedly；Extract liquor rotary evaporation is concentrated with Rotary Evaporators at a temperature of 40 DEG C, it is dense
Contracting liquid final volume is 20mL；Selecting dichloromethane, toxicity is relatively low, and effect of extracting is preferable, is further as extractant
Purifying is prepared；
Lumichrome it is secondarily purified：Do thin-layer chromatography separation to concentrate, thickness of thin layer 0.18mm, by gel into
As instrument observation fluorescence, then concentrate is used column chromatography, the stationary phase of column chromatography is silica gel, and mobile phase is that weight ratio is 6:3:1
N-butanol, methanol, distilled water, obtain the solution of lumichrome after chromatography；By solution in 70 DEG C of rotary evaporations, obtain dense
Contracting liquid, then be placed in vacuum drying chamber and be dried to obtain light yellow lumichrome solid with 55 DEG C；Thin-layer chromatography is suitable for micro-
Amount, volatility are smaller, the separation of substance of not heatproof, and system is distributed in stationary phase and mobile phase according to lumichrome and impurity
The different carry out column chromatography for separation of number, the content of lumichrome is greatly improved after isolating and purifying twice；
The extraction of chlorella protein liquid：By chlorella under the conditions of 2 DEG C, 6000r/min centrifugal concentrating 10 minutes, by concentrate in
Breaking-wall cell is carried out in Ultrasonic Cell Disruptor, cell crushing instrument power is 1.0kW, and probe amplitude is 35%, and it is 30 points to crush the time
Clock, it is broken after gained liquid be placed in 4 DEG C, centrifuge 20 minutes under the conditions of 10000r/min, it is crude protein liquid to take supernatant；It selects super
Sound crushes instrument and crushes chlorella, and shell-broken effect is preferable, and active material will not be impacted, and will not add new pollutant
Matter is conducive to the extraction and purification of chlorella albumen；
The purifying of protein liquid：Protein liquid is placed in saturated ammonium sulfate solution, is chromatographed through DEAE Sepharose Fast Flow
Column, with sodium chloride buffer gradient elution, NaCl concentration gradient 100mmol/L, 200 mmol/L, 400 mmol/L, 700
Mmol/L merges the eluent with enzyme activity, saturating as the phosphate buffer solution that 1.0mM, pH are 7.5 using concentration at a temperature of 2 DEG C
Analysis overnight, obtains protein liquid after purification；With sodium chloride buffer gradient elution, not only separation accuracy is high, and efficiency is higher, and not
New pollutant can be introduced；
The compounding of reagent：50mg lauryl sodium sulfate, 0.5g lumichrome powder are added in protein liquid, stirring is equal
It is even, 30 minutes are stood, adds in stabilizer, stabilizer is 10% glycerine, 2.0mMEDTA, the mixed liquor of 0. 5mMDTT, pH 7.8,
Up to for the reagent for more changing aromatic hydrocarbons selective degradation；It, can be high using photopigment lumichrome as polycyclic aromatic hydrocarbon detection probe
Effect positioning polycyclic aromatic hydrocarbon improves the efficiency for polycyclic aromatic hydrocarbon selective degradation, while the chlorella protein liquid of reagent release can
It is effectively broken for the benzene ring structure of polycyclic aromatic hydrocarbon, is the substances such as aldehyde, alcohol by degrading polycyclic aromatic hydrocarbons, so as to targetedly right
It is completed degradation and eliminates.
Embodiment 2：
A kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation, synthesis, dimethyl including lumichrome
Once purifying, secondarily purified, chlorella protein liquid the extraction of lumichrome, the purifying of protein liquid, the reagent of isoalloxazine
Compounding, specifically include following steps：
The synthesis of lumichrome：6, the 7 dimethyl -9- formic acid ylmethyl isoalloxazines that 1.2g is taken to dry（FMF）Solid is placed in
In beaker, the boric acid solution of 32mL 0.12M and the sodium hydroxide solution of 135mL 0.12M are added in, is protected from light and is stirred to react 100 points
Clock obtains lumichrome, reaction principle such as Fig. 1；It is being protected from light, under alkaline condition, FMF is sloughed aldehyde-base is prepared diformazan
Base isoalloxazine, preparation method is more mild, and reaction process is easily-controllable, simple for process；
The once purifying of lumichrome：Filtrate is placed in separatory funnel by filtering reacting liquid, adds in dichloromethane extraction,
Extraction is until extract liquor is colourless repeatedly；Extract liquor rotary evaporation is concentrated with Rotary Evaporators at a temperature of 38 DEG C, it is dense
Contracting liquid final volume is 18mL；Selecting dichloromethane, toxicity is relatively low, and effect of extracting is preferable, is further as extractant
Purifying is prepared；
Lumichrome it is secondarily purified：Do thin-layer chromatography separation to concentrate, thickness of thin layer 0.2mm, by gel into
As instrument observation fluorescence, then concentrate is used column chromatography, the stationary phase of column chromatography is silica gel, and mobile phase is that weight ratio is 8:5:1
N-butanol, methanol, distilled water, obtain the solution of lumichrome after chromatography；By solution in 72 DEG C of rotary evaporations, obtain dense
Contracting liquid, then be placed in vacuum drying chamber and be dried to obtain light yellow lumichrome solid with 56 DEG C；Thin-layer chromatography is suitable for micro-
Amount, volatility are smaller, the separation of substance of not heatproof, and system is distributed in stationary phase and mobile phase according to lumichrome and impurity
The different carry out column chromatography for separation of number, the content of lumichrome is greatly improved after isolating and purifying twice；
The extraction of chlorella protein liquid：By chlorella under the conditions of 4 DEG C, 8000r/min centrifugal concentrating 10 minutes, by concentrate in
Breaking-wall cell is carried out in Ultrasonic Cell Disruptor, cell crushing instrument power is 1.2kW, and probe amplitude is 38%, and it is 45 points to crush the time
Clock, it is broken after gained liquid be placed in 4 DEG C, centrifuge 20 minutes under the conditions of 12000r/min, it is crude protein liquid to take supernatant；It selects super
Sound crushes instrument and crushes chlorella, and shell-broken effect is preferable, and active material will not be impacted, and will not add new pollutant
Matter is conducive to the extraction and purification of chlorella albumen；
The purifying of protein liquid：Protein liquid is placed in saturated ammonium sulfate solution, is chromatographed through DEAE Sepharose Fast Flow
Column, with sodium chloride buffer gradient elution, NaCl concentration gradient 100mmol/L, 200 mmol/L, 400 mmol/L, 700
Mmol/L merges the eluent with enzyme activity, saturating as the phosphate buffer solution that 1.3mM, pH are 7.6 using concentration at a temperature of 4 DEG C
Analysis overnight, obtains protein liquid after purification；With sodium chloride buffer gradient elution, not only separation accuracy is high, and efficiency is higher, and not
New pollutant can be introduced；
The compounding of reagent：55mg lauryl sodium sulfate, 0.6g lumichrome powder are added in protein liquid, stirring is equal
It is even, 45 minutes are stood, adds in stabilizer, stabilizer is 12% glycerine, 2.3mMEDTA, 0.6mMDTT, 0.8 μM of 3-HPA
Mixed liquor, pH is for 7.6 to get for the reagent for more changing aromatic hydrocarbons selective degradation；3-HPA easily occurs poly- in aqueous solution
Symphysis is into dimer, the aldehyde radical and dithiothreitol (DTT) of dimer（DTT）Hydroxyl between can be connected by hydrogen bond, dimer with
The effect of dithiothreitol (DTT) helps to improve protective effect of the dithiothreitol (DTT) to protein, stablizes so as to improve the storage of reagent
Property and the degradation capability to polycyclic aromatic hydrocarbon, greatly improve the service life of reagent, reduce cost；It is coughed up so that photopigment dimethyl is different
Piperazine is polycyclic aromatic hydrocarbon detection probe, can efficiently position polycyclic aromatic hydrocarbon, improves the efficiency for polycyclic aromatic hydrocarbon selective degradation, simultaneously
The chlorella protein liquid of reagent release can be effectively broken for the benzene ring structure of polycyclic aromatic hydrocarbon, be by degrading polycyclic aromatic hydrocarbons
The substances such as aldehyde, alcohol eliminate so as to targetedly complete degradation to it.
The reagent obtained in embodiment 1,2 is acted on into polycyclic aromatic hydrocarbon and oil, it is as shown in table 1 to calculate its removal rate.
Reagent counts the removal effect of polycyclic aromatic hydrocarbon in 1. embodiment 1,2 of table
As can be seen from Table 1, the embodiment that compares 1, embodiment 2 are more preferable to the removal effect of polycyclic aromatic hydrocarbon and oil content.
Embodiment 3：
Embodiment 3 is differed only in embodiment 1, and lumichrome is synthesized in embodiment 3, and the reaction time is small for 0.5
When.
Embodiment 4：
Embodiment 4 is differed only in embodiment 1, extractant used during the once purifying of lumichrome in embodiment 4
For chloroform.
Embodiment 5：
Embodiment 5 is differed only in embodiment 1, extractant used during the once purifying of lumichrome in embodiment 5
For acetone.
Embodiment 6：
Embodiment 6 is differed only in embodiment 1, in embodiment 6 lumichrome it is secondarily purified when, the stream of column chromatography
Dynamic phase is：N-butanol：Ethyl alcohol：Water=7:2:1.
Embodiment 7：
Embodiment 7 is differed only in embodiment 1, in embodiment 7 lumichrome it is secondarily purified when, the stream of column chromatography
Dynamic phase is：N-butanol：Ethyl alcohol：Water=6:2:2.
Embodiment 8：
Embodiment 8 is differed only in embodiment 1, and in embodiment 8 during small extraction ball algae protein liquid, centrifuge speed is
12000g/min。
Embodiment 9：
Embodiment 9 is differed only in embodiment 1, when ball algae protein liquid is extracted in embodiment 9, centrifuge speed 8000g/
min。
Embodiment 10：
Embodiment 10 is differed only in embodiment 1, the compounding kind of reagent, the addition of lauryl sodium sulfate in embodiment 10
It measures as 80mg.
Embodiment 11：
Embodiment 11 is differed only in embodiment 1, the compounding kind of reagent, the addition of lauryl sodium sulfate in embodiment 11
It measures as 20mg.
The reagent obtained in embodiment 3-11 is acted on into polycyclic aromatic hydrocarbon and oil, it is as shown in table 2 to calculate its removal rate.
Reagent counts the removal effect of polycyclic aromatic hydrocarbon in 2. embodiment 3-11 of table
Routine operation in operating procedure of the present invention is well known to those skilled in the art, herein without repeating.
Technical scheme is described in detail in embodiment described above, it should be understood that the above is only
For specific embodiments of the present invention, it is not intended to limit the invention, all any modifications made in the spirit of the present invention,
Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.
Claims (9)
1. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation, it is characterised in that：The preparation method includes：
The synthesis of lumichrome：By 1.0-1.2g6,7 dimethyl -9- formic acid ylmethyl isoalloxazine solids are placed in beaker,
Boric acid solution and sodium hydroxide solution are added in, is protected from light and is stirred to react 90-100 minutes to obtain lumichrome；
The once purifying of lumichrome：Filtrate is placed in separatory funnel by filtering reacting liquid, adds in dichloromethane extraction,
Extraction is until extract liquor is colourless repeatedly；Extract liquor rotary evaporation is concentrated with Rotary Evaporators at a temperature of 38-40 DEG C,
Concentrate final volume is 18-20mL；
Lumichrome it is secondarily purified：Thin-layer chromatography separation, column chromatography for separation are done to concentrate, dimethyl is obtained after chromatography
The solution of isoalloxazine；By solution in 70-74 DEG C of rotary evaporation, concentrate is obtained, then is placed in dry with 55-58 DEG C in vacuum drying chamber
It is dry to obtain light yellow lumichrome solid；
The extraction of chlorella protein liquid：By chlorella centrifugal concentrating 8-10 minutes under the conditions of 2-4 DEG C, 6000-8000r/min,
Concentrate is subjected to breaking-wall cell in Ultrasonic Cell Disruptor, it is crude protein liquid that centrifugation, which takes supernatant,；
The purifying of protein liquid：Protein liquid is placed in saturated ammonium sulfate solution, is chromatographed through DEAE Sepharose Fast Flow
Column with sodium chloride buffer gradient elution, merges the eluent with enzyme activity, saturating with phosphate buffer solution at a temperature of 0-4 DEG C
Analysis overnight, obtains protein liquid after purification；
The compounding of reagent：Lauryl sodium sulfate, lumichrome powder are added in protein liquid, is stirred evenly, stands 30-
45 minutes, stabilizer was added in get for the reagent for more changing aromatic hydrocarbons selective degradation.
2. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：The concentration of boric acid solution is 0.10-0.12M, volume 30-32mL in the synthesis step of the lumichrome；Hydrogen-oxygen
The concentration for changing sodium solution is 0.1-0.12M, volume 130-135mL.
3. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：In the secondarily purified step of the lumichrome the separated thickness of thin layer of thin-layer chromatography be 0.18-0.25mm, column chromatography
Stationary phase for silica gel, mobile phase is that weight ratio is 6-8:3-5:1 n-butanol, methanol, distilled water.
4. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：In the extraction step of the chlorella protein liquid, cell crushing instrument power is 1.0-1.2kW, and probe amplitude is 35-38%, is broken
The broken time is 30-45 minutes.
5. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：In the extraction step of the chlorella protein liquid, the temperature of extraction is centrifuged as 2-4 DEG C, rotating speed 10000-13000r/min,
Extraction time is 15-20 minutes.
6. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：The concentration gradient of sodium chloride buffer is 0-700mmol/L in the purification step of the protein liquid.
7. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：The concentration of phosphate buffer solution in the purification step of the protein liquid is 1.0-1.3mM, pH 7.4-7.6.
8. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：In the compounding step of the reagent, the additive amount of lauryl sodium sulfate is 50-55mg, and lumichrome powder adds
Dosage is 0.5-0.6g.
9. a kind of preparation method of reagent for polycyclic aromatic hydrocarbon selective degradation according to claim 1, feature exist
In：In the compounding step of the reagent, stabilizer is the mixing of 10-12% glycerine, 2.0-2.3mMEDTA, 0. 5-0.6mMDTT
Liquid, pH 7.6-7.8.
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| CN109096282A (en) * | 2018-08-08 | 2018-12-28 | 浙江海洋大学 | A kind of preparation method of the reagent for sulfa antibiotics selective degradation |
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