CN107632093A - A kind of ultrasonic degradation method of mycotoxin - Google Patents
A kind of ultrasonic degradation method of mycotoxin Download PDFInfo
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- CN107632093A CN107632093A CN201710840986.7A CN201710840986A CN107632093A CN 107632093 A CN107632093 A CN 107632093A CN 201710840986 A CN201710840986 A CN 201710840986A CN 107632093 A CN107632093 A CN 107632093A
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- mycotoxin
- supersound process
- ochratoxin
- aflatoxin
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Abstract
The invention provides a kind of ultrasonic degradation method of mycotoxin, including：A) solution of mixed fungus toxin is provided；The mycotoxin includes two or more in aflatoxin B1, deoxynivalenol enol, zearalenone and ochratoxin；B) the mixed fungus toxin is ultrasonically treated；The time of the supersound process is 10~50min；The amplitude of the supersound process is 10%~50%；The pulse duty factor of the supersound process is 20%~60%.The present invention uses ultrasonic degradation method degrading aflatoxin B 1, deoxynivalenol enol, zearalenone and ochratoxin, with reference to the pulse duty factor of specific sonication treatment time, the amplitude being ultrasonically treated and supersound process, a variety of toxin of degrading simultaneously are allowed the invention to, and degradation rate is high.Biodegrading process of the present invention is simple, and quickly, noresidue is pollution-free.
Description
Technical field
The present invention relates to biological technical field, more particularly, to a kind of ultrasonic degradation method of mycotoxin.
Background technology
Mycotoxin is some fungies, embraces and belongs to such as aspergillus, Penicillium and sickle, caused in growth course easily to cause people
Abnormal secondary metabolite, very high to humans and animals toxicity with animal pathological change and physiology.Existing 300 kinds are found so far very
Verticillium toxin, wherein representative mycotoxin has single-ended Bao Mei alkene race toxin (such as DON), zearalenone (ZEN), volt
Horse toxin Bl (FBI), aflatoxin (AFT), storage aspertoxin A (OTA) and the toxin of T 1 etc..Mycotoxin is naturally-produced
Toxin, be prevalent in the agricultural product such as corn, wheat, barley, paddy, oat, sorghum, flavouring, nut, dairy products and feed
In, the yield of crops is had a strong impact on, the quality of agricultural product is reduced, causes huge economic losses.
After mycotoxin pollutes grain and feed, into food chain, so as to influence breeding performonce fo animals and health of people.By
It is varied in mycotoxin chemistry, biology and toxicologic properties, therefore its toxic action difference is also very big, depending on it is taken the photograph
Enter in level, open-assembly time, animal species, health and feed or food while exist and act synergistically between mycotoxin
Deng.But its common toxicity mainly causes two aspects of DNA damage and cytotoxicity；Specifically, mycotoxin major toxicity is made
With including carcinogenesis, genetoxic, teratogenesis, hepatotoxicity, nephrotoxicity, genital disorders and immunosupress.
According to FAO (Food and Agriculture Organization of the United Nation) (FAO) report, the crops that the whole world there are about 25% every year are polluted by fungi and toxin, there are about
2% crops lose nutrition and economic value because seriously polluted, caused by directly or indirectly economic loss reach tens billion of U.S.s
Member.Both at home and abroad to the aflatoxin in Cereals, food and feed, ochratoxin A, deoxynivalenol, corn
The mycotoxins such as red mould dilute ketone have formulated strict limit standard.
The method effect of the disclosed processing mycotoxin of prior art is preferable not enough, mainly including Physical, chemical method,
Absorption method, bioanalysis etc..Physical detoxification is not thorough, and Environmental costs are high, and is mostly directed to the processing of single toxin；Chemical method
It is quick rapid, but noxious material is easily remained, influence quality；Absorption method needs to carry out desorption processing, is also easy to produce secondary pollution；It is raw
The processing of thing method is time-consuming longer, and cost is too high, and metabolite toxicity is unstable.
The content of the invention
In view of this, the technical problem to be solved in the present invention is to provide a kind of ultrasonic degradation method of mycotoxin, sheet
The biodegrading process of invention is simple, and can degrade a variety of toxin simultaneously, and degradation rate is high.
The invention provides a kind of ultrasonic degradation method of mycotoxin, including：
A the solution that mixed fungus toxin is provided) is prepared；The mycotoxin includes aflatoxin B1, rotten sickle is avenged in deoxidation
Two or more in knife enol, zearalenone and ochratoxin；
B) the mixed fungus toxin is ultrasonically treated；The time of the supersound process is 10~50min；It is described super
The amplitude of sonication is 10%~50%；The pulse duty factor of the supersound process is 20%~60%.
Preferably, step A) solution of the mycotoxin is the aqueous solution.
Preferably, the mycotoxin includes aflatoxin B1, deoxynivalenol enol, zearalenone and reddish brown
Aspertoxin；Aflatoxin B1, deoxynivalenol enol, zearalenone and reddish brown song in the solution of the mycotoxin
The concentration ratio of mould toxin is：1~10：50~100:1~20:1~10.
Preferably, step B) ultrasonic system is using probe type ultrasonic system；The probe gos deep into below liquid level
Depth be 0.5~2.0cm.
Preferably, step B) supersonic frequency is 20000Hz；Input power is 100~550W.
Preferably, step B) time of the supersound process is 10~40min；The amplitude of the supersound process be 20%~
50%；The pulse duty factor of the supersound process is 20%~50%.
Preferably, step B) it is described supersound process complete cycle be specially：1~5s of ultrasound, pause 1~10s.
The invention provides a kind of High Performance Liquid Chromatography/Mass Spectrometry detection method of mycotoxin, including：
Mixture mycotoxin is dissolved in methanol aqueous solution, obtains prepare liquid；
Prepare liquid is subjected to High Performance Liquid Chromatography/Mass Spectrometry detection, then compareed with the testing result of standard items prepare liquid,
Obtain quantitative data；
The chromatographic condition is：Chromatographic column：C18；Mobile phase A：Methanol；B：Ammonium acetate-aqueous formic acid；Flow velocity 300ul/
min；Condition of gradient elution：0~1.5min, the volume fraction of mobile phase A rise to 95% by 25%；1.5~4.2min, mobile phase
A volume fraction is maintained at 95%；The volume fraction of 4.2~4.3min mobile phase As is down to 25% by 95%；4.3~6min flows
Dynamic phase A volume fraction is maintained at 25%；
Mass Spectrometry Conditions：Ionization mode is ESI, and spray voltage is 3600V, and polar mode is cation, and heating-up temperature is
260 DEG C~270 DEG C, sheath gas is 40arb, and auxiliary gas is 12arb, and capillary temperature is 320 DEG C~330 DEG C.
Preferably, the sample size is 5~6uL.
Preferably, the compound method of the mycotoxin standard items prepare liquid is specially：Mycotoxin standard items are used
Acetonitrile dissolving, nitrogen blow, then are redissolved using 25% methanol aqueous solution.
Compared with prior art, the invention provides a kind of ultrasonic degradation method of mycotoxin, including：A) provide mixed
Close the solution of mycotoxin；The mycotoxin include aflatoxin B1, deoxynivalenol enol, zearalenone and
Two or more in ochratoxin；B) the mixed fungus toxin is ultrasonically treated；The supersound process
Time is 10~50min；The amplitude of the supersound process is 10%~50%；The pulse duty factor of the supersound process is 20%
~60%.The present invention is using ultrasonic degradation method degrading aflatoxin B 1, deoxynivalenol enol, zearalenone
And ochratoxin, with reference to the pulse duty factor of specific sonication treatment time, the amplitude being ultrasonically treated and supersound process, make
A variety of toxin of the invention that can degrade simultaneously are obtained, and degradation rate is high.Biodegrading process of the present invention is simple, quickly, noresidue, no dirt
Dye.
Brief description of the drawings
Fig. 1 is the extraction ion flow chromatography of the blank sample of the embodiment of the present invention 5；
Fig. 2 be the ultrasonic degradation of the embodiment of the present invention 5 after sample extraction ion stream chromatogram.
Embodiment
The invention provides a kind of ultrasonic degradation method of mycotoxin, including：
A) solution of mixed fungus toxin is provided；The mycotoxin includes aflatoxin B1, deoxynivalenol alkene
Two or more in alcohol, zearalenone and ochratoxin；
B) the mixed fungus toxin is ultrasonically treated；The time of the supersound process is 10~50min；It is described super
The amplitude of sonication is 10%~50%；The pulse duty factor of the supersound process is 20%~60%.
The ultrasonic degradation method of mycotoxin provided by the invention is applied to degrade while a variety of toxin.
Ultrasonic wave is the characteristics such as a kind of frequency is high, wavelength is small, penetration power is strong, and frequency exceedes 20kHz sound wave.Ultrasonic wave
With the interphase interaction of medium, cavitation effect is produced.Cavitation effect just refers to that the micro-bubble in fluid is formed, grown, be broken
The phenomenon split, the rupture of cavitation bubble can locally produce the focus of high temperature (about 5000K) high pressure (about 5000atm) in bubble, also
There are heating rapidly and cooling (being more than 109K/sec) and a liquid micro-jet (about 400km/h), and the focus of this HTHP
Can greatly accelerates the chemical reaction in medium.Ultrasonic technology mainly has destruction to the biochemical characteristic and physical characteristic of food
Inductive effect.The technology repair by the crystalline texture modification (sound crystallization) applied to modified fat, the structure of different food proteins
Decorations etc..
Present invention firstly provides the solution of mixed fungus toxin；The mycotoxin includes aflatoxin B1, deoxidation is avenged
Two or more in rotten reaping hook enol, zearalenone and ochratoxin；Preferably include aflatoxin B1, take off
Three kinds or more in oxygen deoxynivalenol, zearalenone and ochratoxin；More preferably include aspergillus flavus poison
Plain B1, deoxynivalenol enol, zearalenone and ochratoxin.As：Rotten sickle is avenged in aflatoxin AFB1, deoxidation
Knife enol DON, zearalenone ZEN, ochratoxin OTA.
According to the present invention, the solution of the mycotoxin is the aqueous solution.Aflatoxin in the solution of the mycotoxin
B1, deoxynivalenol enol, the concentration ratio of zearalenone and ochratoxin are preferably：1~10：50~100:1~
20:1~10；More preferably 3~10：60~100:5~15:1~5；Most preferably 4：100:10:1.
The mixed fungus toxin is ultrasonically treated.
For the present invention for the ultrasonic instrument without limiting, those skilled in the art are known；It is preferred that can be
BRANSON SFX550 ultrasonic equipments, standard frequency are preferably 20000Hz, and input power is preferably 100~550W；More preferably
200~550W, most preferably 300~~550W；Amplitude adjustable range 10%~70%.
In the present invention, the ultrasonic system is preferably using probe type ultrasonic system；It is described probe go deep into liquid level with
Under depth be preferably 0.5~2.0cm；More preferably 1.5cm~2.0cm；Most preferably 1.5cm.
The time of the supersound process is 10~50min；Preferably 10~40min；The amplitude of the supersound process is
10%~50%；Preferably 20%~50%；The pulse duty factor of the supersound process is 20%~60%；Preferably 20%~
50%.
Sonication treatment time of the present invention is the ultrasound works time.
The pulse duty factor of supersound process of the present invention refers to the ratio of a circulation shared by the time of supersound process, than
Paused 2 seconds within 2 seconds such as ultrasound, pulse duty factor 50%.
In the present invention, the supersound process complete cycle is preferably specially：1~5s of ultrasound, pause 1~10s；More preferably
Specially 1~2s of ultrasound, pause 1~2s.
Pulse ultrasonic processing method time bag ultrasonic time and interval time in ultrasonic applications, this is one complete
Sonication cycles time, the time range of the present invention preferably one circulation of the ultrasound completion 2~15 seconds.
After supersound process, mycotoxin is degraded.
The invention provides a kind of ultrasonic degradation method of mycotoxin, including：A) the molten of mixed fungus toxin is provided
Liquid；The mycotoxin is included in aflatoxin B1, deoxynivalenol enol, zearalenone and ochratoxin
Two or more；B) the mixed fungus toxin is ultrasonically treated；The time of the supersound process be 10~
50min；The amplitude of the supersound process is 10%~50%；The pulse duty factor of the supersound process is 20%~60%.This
Invention uses ultrasonic degradation method degrading aflatoxin B 1, deoxynivalenol enol, zearalenone and Aspergillus ochraceus
Toxin, with reference to the pulse duty factor of specific sonication treatment time, the amplitude being ultrasonically treated and supersound process so that the present invention
Can be degraded a variety of toxin simultaneously, and degradation rate is high.Biodegrading process of the present invention is simple, and quickly, noresidue is pollution-free.
After supersound process, preferably through cooling, nitrogen blows, and 25% methanol redissolves, and is detected.
The invention provides a kind of High Performance Liquid Chromatography/Mass Spectrometry detection method of mycotoxin, including：
Mixture mycotoxin is dissolved in methanol aqueous solution, obtains prepare liquid；
Prepare liquid is subjected to High Performance Liquid Chromatography/Mass Spectrometry detection, then compareed with the testing result of standard items prepare liquid,
Obtain quantitative data；
The chromatographic condition is：Chromatographic column：C18；Mobile phase A：Methanol；B：Ammonium acetate-aqueous formic acid；Flow velocity 300ul/
min；Condition of gradient elution：0~1.5min, the volume fraction of mobile phase A rise to 95% by 25%；1.5~4.2min, mobile phase
A volume fraction is maintained at 95%；The volume fraction of 4.2~4.3min mobile phase As is down to 25% by 95%；4.3~6min flows
Dynamic phase A volume fraction is maintained at 25%；
Mass Spectrometry Conditions：Ionization mode is ESI, and spray voltage is 3600V, and polar mode is cation, and heating-up temperature is
260 DEG C~270 DEG C, sheath gas is 40arb, and auxiliary gas is 12arb, and capillary temperature is 320 DEG C~330 DEG C.
The High Performance Liquid Chromatography/Mass Spectrometry detection method of mycotoxin provided by the invention is molten by mixture mycotoxin first
In methanol aqueous solution, prepare liquid is obtained.
Said mixture fungi can think original testing mixture fungi or the ultrasonotomography using the present invention
Mixture fungi after method degraded.
It is preferably specially that mixture mycotoxin is dissolved in into 25% that mixture mycotoxin, which is dissolved in methanol aqueous solution,
In methanol aqueous solution, prepare liquid is obtained.
Mixture mycotoxin standard liquid can also be configured simultaneously, obtains standard items prepare liquid.
The configuration mode is preferably:The offer method of the mycotoxin standard items prepare liquid is specially：By fungi poison
Plain standard items are blown using acetonitrile dissolving, nitrogen, then are redissolved using 25% methanol aqueous solution.
More preferably it is specially：
Four kinds of mycotoxin standard sample mother liquors are first prepared respectively, and four kinds of mycotoxins are dissolved with acetonitrile, and concentration is
100ppm (0.1mg/ml), in -20 DEG C of storages.Prepare the middle interstitial fluid of four kinds of mycotoxins, DON respectively with pure dilution in acetonitrile again
10ppm, AFB1 1ppm, ZEN 1ppm, OTA 1ppm, in 4 DEG C of refrigerator storages.Afterwards according to DON:ZEN:AFB1:OTA=
100:10:4:1 ratio prepares the mixing standard liquid of four kinds of mycotoxins,
The concentration of each mycotoxin is as shown in table 1.The mixing standard liquid prepared needs nitrogen to blow, then with isometric 25% methanol
The aqueous solution redissolves, then enters liquid chromatogram and carry out analysis detection, draws quantitation curves.
The concentration of the mixed fungus toxin standard items of the quantitation curves of table 1
Prepare liquid is subjected to High Performance Liquid Chromatography/Mass Spectrometry detection, then compareed with the testing result of standard items prepare liquid,
Obtain quantitative data.
Present invention preferably employs be Thermo liquid chromatographic systems.
The chromatographic condition is preferably specially：
Chromatographic column：C18；
Mobile phase A：Methanol；B：Ammonium acetate-aqueous formic acid；
Flow velocity 300ul/min；The sample size is 5uL.
Condition of gradient elution：0~1.5min, the volume fraction of mobile phase A rise to 95% by 25%；1.5~4.2min, stream
Dynamic phase A volume fraction is maintained at 95%；The volume fraction of 4.2~4.3min mobile phase As is down to 25% by 95%；4.3~
The volume fraction of 6min mobile phase As is maintained at 25%.
The Mass Spectrometry Conditions are preferably specially：
Ionization mode is ESI,
Spray voltage is 3600V,
Polar mode is cation,
Heating-up temperature is 260 DEG C~270 DEG C,
Sheath gas is 40arb, and auxiliary gas is 12arb,
Capillary temperature is 320 DEG C~330 DEG C.
Four kinds of mycotoxins can be detected simultaneously, examined using High Performance Liquid Chromatography/Mass Spectrometry detection method of the present invention
Survey method is simple, and preci-sion and accuracy is high, and detection limit is low.
In order to further illustrate the present invention, smoke composition provided by the invention is retouched in detail with reference to embodiments
State.
Embodiment 1
First prepare four kinds of mycotoxin (aflatoxin AFB respectively according to the method described in the present invention1, deoxidation avenge rotten sickle
Knife enol DON, zearalenone ZEN, ochratoxin OTA) standard sample mother liquor, four kinds of mycotoxins dissolve with acetonitrile,
Concentration is 100ppm (0.1mg/ml), in -20 DEG C of storages.Prepare the centre of four kinds of mycotoxins respectively with pure dilution in acetonitrile again
Liquid, DON 10ppm, AFB1 1ppm, ZEN 1ppm, OTA 1ppm, in 4 DEG C of refrigerator storages.Afterwards according to DON:ZEN:AFB1:
OTA=100:10:4:1 ratio prepares the mixing standard liquid of four kinds of mycotoxins, and the concentration of each mycotoxin is as shown in table 1.Institute
The mixing standard liquid of preparation needs nitrogen to blow, then is redissolved with isometric 25% methanol aqueous solution, then enters liquid chromatogram and carry out analysis detection,
Draw quantitation curves.
Using Thermo liquid chromatographic systems, using Thermo UHPLC/ESI Q-Orbitrap mass spectrometer systems, chromatostrip
Part is：Chromatographic column：C18；Mobile phase A：Methanol；B：Ammonium acetate-aqueous formic acid；Flow velocity 300ul/min；Condition of gradient elution：0
~1.5min, the volume fraction of mobile phase A rise to 95% by 25%；1.5~4.2min, the volume fraction of mobile phase A are maintained at
95%；4.2 the volume fraction of~4.3min mobile phase As is down to 25% by 95%；The volume fraction of 4.3~6min mobile phase As is protected
Hold 25%；
Mass Spectrometry Conditions：Ionization mode is ESI, and spray voltage is 3600V, and polar mode is cation, and heating-up temperature is
260 DEG C, sheath gas is 40arb, and auxiliary gas is 12arb, and capillary temperature is 320 DEG C.
As a result show, the standard curve of four kinds of toxin is as follows：
AFB1：Y=616466+1.63787e+006X R2=0.9971；
DON:Y=18402+1.94279e+006X R2=0.9973；
ZEN:Y=-39861.15+71365.8X R2=0.9966；
OTA:Y=-3290.45+107908X R2=0.9963.
The precision of embodiment 2, recovery of standard addition experiment
The standard items solvent of four kinds of mycotoxins is subjected to withinday precision n=10 and day to day precision experiment n=5, knot
Fruit is as shown in table 2 and table 3, and wherein table 2 is that the standard items solvent of four kinds of mycotoxins carries out withinday precision result, and table 3 is four
The standard items solvent of kind mycotoxin carries out day to day precision.
The standard items solvent of 2 four kinds of mycotoxins of table carries out withinday precision result
The standard items solvent of 3 four kinds of mycotoxins of table carries out day to day precision result
Meanwhile the sensitivity of the method assay method by standard solution gradient dilution of the inventive method, according to RSN
≥10；
The quantitative limit scope of method is 1~50 μ g/kg, and detection is limited to 0.5~25 μ g/kg；
The rate of recovery of method is investigated using mark-on method, the TIANZHU XINGNAO Capsul of high, medium and low parallel 3 parts of three concentration
Scope is 85.7%~109.0%.
Embodiment 3
Prepare four kinds of mycotoxin (aflatoxin AFB1, deoxynivalenol enol DON, zearalenone ZEN,
Ochratoxin OTA) standard items mixed aqueous solution, its concentration is respectively DON 2000ppb, AFB1 80ppb,ZEN
200ppb,OTA 20ppb.Take mixing mark product aqueous solution 25ml that ultrasonic probe is entered 1.5cm in sample in plastics small beaker
Deep to carry out ultrasonication, ultrasonic instrument condition is fixed frequency 20000Hz, input amplitude 20%, ultrasonic time 10 minutes,
Pulse duty factor be 33.3% be ultrasound 1 second, pause 2 seconds be a complete ultrasound processing cycle.Sample after processing etc.
After cooling, take 2ml nitrogen to blow, then dissolved with 25% methanol-water, enter liquid phase and detected, parallel three samples.According to mark song and sky
The ratio of white sample draws aflatoxin AFB1Degradation rate is 58.1%, deoxynivalenol enol DON degradation rates be 18.8%,
Zearalenone ZEN degradation rates are 66.4%, ochratoxin OTA degradation rates are 51.4%.
Embodiment 4
Prepare four kinds of mycotoxin (aflatoxin AFB1, deoxynivalenol enol DON, zearalenone ZEN,
Ochratoxin OTA) the standard items aqueous solution, its concentration is respectively DON 500ppb, AFB1 20ppb,ZEN 50ppb,OTA
5ppb.Mixing mark product aqueous solution 25ml is taken in plastics small beaker, ultrasonic probe is entered 1.5cm in sample carries out ultrasonic wave deeply
Processing, ultrasonic instrument condition are fixed frequency 20000Hz, input amplitude 50%, ultrasonic time 40 minutes, and pulse duty factor is
Pulse duty factor be 33.3% be ultrasound 1 second, pause 2 seconds be a complete ultrasound processing cycle.Sample after processing etc.
After cooling, take 2ml nitrogen to blow, then dissolved with 25% methanol-water, enter liquid phase and detected, parallel three samples.According to mark song and sky
The ratio of white sample draws aflatoxin AFB1Degradation rate is 93.6%, deoxynivalenol enol DON degradation rates be 53.0%,
Zearalenone ZEN degradation rates are 95.0%, ochratoxin OTA degradation rates are 77.2%.
Embodiment 5
Prepare four kinds of mycotoxin (aflatoxin AFB1, deoxynivalenol enol DON, zearalenone ZEN,
Ochratoxin OTA) the standard items aqueous solution, its concentration is respectively DON 500ppb, AFB1 20ppb,ZEN 50ppb,OTA
5ppb.Mixing mark product aqueous solution 25ml is taken in plastics small beaker, ultrasonic probe is entered 1.5cm in sample carries out ultrasonic wave deeply
Processing, ultrasonic instrument condition are fixed frequency 20000Hz, input amplitude 20%, ultrasonic time 40 minutes, and pulse duty factor is
25% be ultrasound 1 second, pause 3 seconds be a complete ultrasound processing cycle.After the cooling such as sample after processing, 2ml nitrogen is taken
Blow, then dissolved with 25% methanol-water, enter liquid phase and detected, parallel three samples.Fig. 1 is the blank sample of the embodiment of the present invention 5
Extract ion flow chromatography；Fig. 2 be the ultrasonic degradation of the embodiment of the present invention 5 after sample extraction ion stream chromatogram.According to mark
Bent and blank sample ratio draws aflatoxin AFB1Degradation rate is 96.5%, and deoxynivalenol enol DON degradation rates are
61.5%th, zearalenone ZEN degradation rates be 96.9%, ochratoxin OTA degradation rates be 91.6%.
Comparative example 1
Prepare four kinds of mycotoxin (aflatoxin AFB1, deoxynivalenol enol DON, zearalenone ZEN,
Ochratoxin OTA) standard items mixed aqueous solution, its concentration is respectively DON 2000ppb, AFB1 80ppb,ZEN
200ppb,OTA20ppb.Take mixing mark product aqueous solution 25ml that ultrasonic probe is entered 2.5cm in sample in plastics small beaker
Deep to carry out ultrasonication, ultrasonic instrument condition is fixed frequency 20000Hz, input amplitude 60%, ultrasonic time 5 minutes,
Pulse duty factor be 10% be ultrasound 1 second, pause 9 seconds be a complete ultrasound processing cycle.Sample after processing etc. drops
Wen Hou, take 2ml nitrogen to blow, then dissolved with 25% methanol-water, enter liquid phase and detected, parallel three samples.According to mark song and blank
The ratio of sample draws aflatoxin AFB1Degradation rate is 10.4%, and deoxynivalenol enol DON degradation rates are 2.1%, jade
Zearlenone ZEN degradation rates are 13.5%, ochratoxin OTA degradation rates are 7.4%.
Comparative example 2
Prepare four kinds of mycotoxin (aflatoxin AFB1, deoxynivalenol enol DON, zearalenone ZEN,
Ochratoxin OTA) the standard items aqueous solution, its concentration is respectively DON 500ppb, AFB1 20ppb,ZEN 50ppb,OTA
5ppb.Mixing mark product aqueous solution 25ml is taken in plastics small beaker, ultrasonic probe is entered 0.5cm in sample carries out ultrasonic wave deeply
Processing, ultrasonic instrument condition are fixed frequency 20000Hz, input amplitude 5%, ultrasonic time 60 minutes, and pulse duty factor is
Pulse duty factor be 70% be ultrasound 7 seconds, pause 3 seconds be a complete ultrasound processing cycle.Sample after processing etc. drops
Wen Hou, take 2ml nitrogen to blow, then dissolved with 25% methanol-water, enter liquid phase and detected, parallel three samples.According to mark song and blank
The ratio of sample draws aflatoxin AFB1Degradation rate is 28.1%, and deoxynivalenol enol DON degradation rates are 15.4%, jade
Zearlenone ZEN degradation rates are 40.9%, ochratoxin OTA degradation rates are 34.7%.
Comparative example 3
Prepare four kinds of mycotoxin (aflatoxin AFB1, deoxynivalenol enol DON, zearalenone ZEN,
Ochratoxin OTA) the standard items aqueous solution, its concentration is respectively DON 500ppb, AFB1 20ppb,ZEN 50ppb,OTA
5ppb.Mixing mark product aqueous solution 50ml is taken sample to be placed in plastics small beaker in groove type ultrasonic ripple instrument, ultrasonic instrument
Condition is fixed frequency 40000Hz, input power 500W, input amplitude 100%, continuous ultrasound 25 minutes.After processing
After the cooling such as sample, take 2ml nitrogen to blow, then dissolved with 25% methanol-water, enter liquid phase and detected, parallel three samples.According to mark
Bent and blank sample ratio draws aflatoxin AFB1Degradation rate is 5.1%, and deoxynivalenol enol DON degradation rates are
0.5%th, zearalenone ZEN degradation rates be 7.5%, ochratoxin OTA degradation rates be 3.4%.
Embodiment 6
Ultrasonic wave of the standard items mixed solution of the various concentrations of four kinds of toxin prepared by embodiment 1 described in through embodiment 1
After processing, the content of contratoxin is detected, and mixed standard solution of each toxin without ultrasonication is as control sample, respectively
The degradation rate of toxin is shown in Table 4.
The degradation rate of toxin of the 4 four kinds of mycotoxin mixed standard solutions of table after ultrasonication
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of ultrasonic degradation method of mycotoxin, it is characterised in that including：
A) solution of mixed fungus toxin is provided；The mycotoxin includes aflatoxin B1, deoxynivalenol enol, jade
Two or more in Zearlenone and ochratoxin；
B) the mixed fungus toxin is ultrasonically treated；The time of the supersound process is 10~50min；At the ultrasound
The amplitude of reason is 10%~50%；The pulse duty factor of the supersound process is 20%~60%.
2. biodegrading process according to claim 1, it is characterised in that step A) solution of the mycotoxin is water-soluble
Liquid.
3. biodegrading process according to claim 1, it is characterised in that the mycotoxin includes aflatoxin B1, taken off
Oxygen deoxynivalenol, zearalenone and ochratoxin；Aflatoxin B1, deoxidation in the solution of the mycotoxin
The concentration ratio of deoxynivalenol, zearalenone and ochratoxin is：1~10：50~100:1~20:1~10.
4. biodegrading process according to claim 1, it is characterised in that step B) ultrasonic system is using sonde-type
Ultrasonic system；The depth goed deep into below liquid level of popping one's head in is 0.5~2.0cm.
5. biodegrading process according to claim 1, it is characterised in that step B) supersonic frequency is 20000Hz；Input
Power is 100~550W.
6. biodegrading process according to claim 1, it is characterised in that step B) supersound process time for 10~
40min；The amplitude of the supersound process is 20%~50%；The pulse duty factor of the supersound process is 20%~50%.
7. biodegrading process according to claim 1, it is characterised in that step B) it is described supersound process complete cycle it is specific
For：1~5s of ultrasound, pause 1~10s.
A kind of 8. High Performance Liquid Chromatography/Mass Spectrometry detection method of mycotoxin, it is characterised in that including：
Mixture mycotoxin is dissolved in methanol aqueous solution, obtains prepare liquid；
Prepare liquid is subjected to High Performance Liquid Chromatography/Mass Spectrometry detection, then compareed with the testing result of standard items prepare liquid, is obtained
Quantitative data；
The chromatographic condition is：Chromatographic column：C18；Mobile phase A：Methanol；B：Ammonium acetate-aqueous formic acid；Flow velocity 300ul/min；
Condition of gradient elution：0~1.5min, the volume fraction of mobile phase A rise to 95% by 25%；1.5~4.2min, mobile phase A
Volume fraction is maintained at 95%；The volume fraction of 4.2~4.3min mobile phase As is down to 25% by 95%；4.3~6min mobile phases
A volume fraction is maintained at 25%；
Mass Spectrometry Conditions：Ionization mode is ESI, and spray voltage is 3600V, and polar mode is cation, and heating-up temperature is 260 DEG C
~270 DEG C, sheath gas is 40arb, and auxiliary gas is 12arb, and capillary temperature is 320 DEG C~330 DEG C.
9. detection method according to claim 8, it is characterised in that the sample size is 5~6uL.
10. detection method according to claim 8, it is characterised in that the preparation of the mycotoxin standard items prepare liquid
Method is specially：Mycotoxin standard items are blown using acetonitrile dissolving, nitrogen, then redissolved using 25% methanol aqueous solution.
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