CN110108823A - Organic toxin sample vibration purification detection method and its ultrasonic liquid-transfering gun in a kind of cereal crops - Google Patents
Organic toxin sample vibration purification detection method and its ultrasonic liquid-transfering gun in a kind of cereal crops Download PDFInfo
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- CN110108823A CN110108823A CN201910399966.XA CN201910399966A CN110108823A CN 110108823 A CN110108823 A CN 110108823A CN 201910399966 A CN201910399966 A CN 201910399966A CN 110108823 A CN110108823 A CN 110108823A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
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- G01N30/14—Preparation by elimination of some components
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
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- G01N2030/062—Preparation extracting sample from raw material
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Abstract
The present invention provides organic toxin sample purification extraction detection method in a kind of cereal crops and its ultrasonic liquid-transfering guns, belong to chemical analysis detection technique field, and detecting step includes: sample extraction, sample purification, vibration extraction, sample drain；Pipetting technique is combined with sample purification technology, the purification process for completing sample to be tested while liquid pipettes may be implemented, operating procedure is greatly saved, ensure that detection accuracy.
Description
Technical field
The invention belongs to chemical analysis detection technique fields, and in particular to organic toxin sample vibration in a kind of cereal crops
Purify detection method and its ultrasonic liquid-transfering gun.
Background technique
Cereal crops especially cereals crop, from initial artificial growth or mechanical harvesting, to dry decladding and it is final
It puts in storage, needs to undergo very long storage process.The storage of common grain, does not often add any anti-corrosion in cereal crops
Ingredient depends merely on drying and dehydrating storage.Therefore, grain mildew usually occurs because processing storage is improper or circulation overlong time.
Organic toxin in grain is to be metabolized generation, such as zearalenone, vomitoxin, aspergillus flavus by various moulds
Element etc. is the carcinogenic substance of a variety of severe toxicity.The very important step of grain processing preamble technique is exactly toxic component analysis, before analysis
It needs to carry out purification extraction processing to the grain substance of interference.Common cleaning grains method mostly use water purification cleaning, bubble with
And the modes such as boiling sterilization, then using corresponding organic solvent Solid Phase Extraction.
However, 1. the above method needs special grain processing equipment, and it is at high cost, it is unfavorable for quickly detection and field acquisition
The environment of detection.2. sample only leans on gravity and silica gel absorption to act on and slowly flows through solid phase column in purification process, take long time and
It is inconvenient, and plant material particle is easy blocking cylinder, influences to purify yield.3. sample needs additionally to walk after cleaning
Suddenly it is transferred in detection device and is detected.It is difficult to avoid that is in contact with external environment, easily causes secondary pollution, shadow
Ring the precision of detection.Liquid-transfering gun is that the liquid of analyte detection process indispensability pipettes tool, and liquid relief precision is high, is widely used in grain
Food, environment, field of biological detection.
Summary of the invention
In order to improve detection working efficiency, detection accuracy is improved, the present invention provides organic toxin sample in a kind of cereal crops
Product purification extraction detection method, pipetting technique, sample purification technology and ultrasonic extraction technology are combined, may be implemented in liquid
While body pipettes, the ultrasonic extraction process of sample to be tested is completed, operating procedure is greatly saved, ensure that detection accuracy.
The present invention provides organic toxin sample purification extraction detection method in a kind of cereal crops, and detecting step includes: (1)
Sample extraction: taking measuring samples Extraction solvent constant volume, mechanical shaking extraction, and collection supernatant is sample extracting solution；(2) sample is net
Change: pipette samples extracting solution quantify using liquid-transfering gun, make extracting solution flow through/flow through scavenging material, continue quantitative draw air, it is bulging
Bubble in dynamic sample extracting solution, makes scavenging material adsorbing contaminant in liquid transfer gun head；(3) it vibration extraction: is shaken using liquid-transfering gun ultrasound
Dynamic sample extracting solution is crushed bubble in scavenging material hole with sample extracting solution and mixes, makes scavenging material in gas-liquid interface
Further extracting impurities；(3) sample drain: purified sample is discharged, and is sent to machine testing on detection device；
Wherein, in step (2), sample purification step includes: 1. to take out pipette tips, is installed on liquid-transfering gun, and pipette tips are protruded into
Sample liquid level to be clean makes liquid-transfering gun generate negative-pressure liquid suction in pipette tips hereinafter, press liquid-transfering gun, sucks 1ml sample to be clean,
So that purification sample is flowed through/flow through purification sample, stops 5s；2. liquid-transfering gun pipette tips are removed liquid level, liquid-transfering gun is pressed again, makes to move
Negative-pressure liquid suction is generated in liquid rifle in pipette tips, 1ml air is sucked, agitates bubble in sample extracting solution, fill liquid with scavenging material
Divide mixing, repeat this step three times, sucks 4ml air liquid-transfering gun altogether and show that 5000ul ALL is fully loaded with；3. opening liquid-transfering gun ultrasound
4. pipette tips are directed at 2ml sample bottle, press liquid-transfering gun by energy converter, sample extracting solution and scavenging material in ultrasonic vibration liquid-transfering gun
Drain, machine analysis on collection and purification liquid.
Wherein, in step (1), extraction step includes: to weigh 5g cereal crops sample comminution, crosses 1mm sub-sieve, in
In 50mL centrifuge tube, disperses constant volume to 20mL with extracting solution (95wt% acetonitrile/water), be vortexed and mix；Ultrasonic extraction 30min, puts
Enter centrifuge (8000r/min is centrifuged 20min), takes supernatant as sample extracting solution.The cereal crops are preferably plant
Cereal crops, including corn, wheat and rice；Organic toxin includes zearalenone, patulin and vomitoxin.
Wherein, it in step (3), is analyzed by high performance liquid chromatography (HPLC), zearalenone testing conditions
Are as follows:
Mobile phase: A phase, water；B phase, acetonitrile-methanol-water (46:8:46, v:v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 1.0mL/min；
Column temperature: 35 DEG C；
Sample volume: 50 μ L；
Fluorescence detection: excitation wavelength: 274nm；Launch wavelength: 440nm, photochemistry post-column derivation.
Wherein, it in step (3), is analyzed by high performance liquid chromatography (HPLC-LC-20A), patulin detector bar
Part are as follows:
Mobile phase: A phase, acetonitrile；B phase, acetonitrile -0.5mM ammonium acetate (14:86, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 0.7mL/min；
Column temperature: 30 DEG C；
Sample volume: 20 μ L；
Fluorescence detection: excitation wavelength: 276nm；Launch wavelength: 440nm, photochemistry post-column derivation.
Wherein, it in step (3), is analyzed by high performance liquid chromatography (HPLC-LC-20A), vomitoxin detector bar
Part are as follows:
Mobile phase: methanol-water (20:80, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 1.0mL/min；
Column temperature: 35 DEG C；
Sample volume: 50 μ L；
Ultraviolet detection: excitation wavelength: 218nm.
Scavenging material includes but is not limited to: diatomite, magnesium sulfate, ketjenblack EC, sulfonic group n-vinyl pyrrolidone-
Divinylbenzene, carboxyl n-vinyl pyrrolidone-divinylbenzene, methylene piperazine ring n-vinyl pyrrolidone-diethyl
Alkenyl benzene, C18Modified silica-gel, C8Modified silica-gel, aminopropyl bonded silica gel, glycol-based bonded silica gel, florisil silica or aluminium oxide
At least one of.
The present invention also provides a kind of purifications suitable for above-mentioned detection method to extract liquid-transfering gun comprising gun body, gun body packet
Include handle, ultrasonic transducer, imbibition key, drain key, pipette tips interface and battery pack, ultrasonic transducer ultrasonic probe and pipette tips
Interface is connected, and ultrasonic transducer is electrically connected with imbibition key, drain key and battery pack respectively, and ultrasonic transducer is preferably adopted
With 28khz60W ultrasonic vibrator.
Beneficial technical effect
Organic toxin sample purification provided by the invention extracts detection method, by micropipette technology and sample adsorption cleaning
Technology combines, and may be implemented, by Vltrasonic device in liquid-transfering gun, scavenging material to be made to exist while trace liquid quantitative pipettes
Gas-liquid interface between extracting solution and bubble, fast and effective extracting impurities ingredient, and agitated by the shatter micro-bubble of ultrasound, promote
It is sufficiently mixed absorption into scavenging material, completes the purification process of sample to be tested, operating procedure is greatly saved, eliminates impurity interference
Background ensure that detection accuracy, and under the premise of guaranteeing sample recovery rate, improve sample purification speed, detection repeatability
It is good.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of ultrasound liquid-transfering gun used in organic toxin of the present invention purification extraction detection method；
Fig. 2, which is that organic toxin of the present invention purification extraction detection method is used, purifies pipette tips pipette samples schematic diagram；
Fig. 3 is that organic toxin of the present invention purification extraction detection method purification pipette tips used purify sample schematic diagram.
Fig. 4 is that organic toxin of the present invention purification extraction detection method purification pipette tips used discharge sample schematic diagram.
Fig. 5 is that organic toxin of the present invention purification extraction detection method embodiment detects spectrogram:
The mixed mark liquid chromatogram of 1 Fig. 5 a organic toxin of embodiment；Fig. 5 b maize flour negative sample and negative mark-on sample pair
Than figure；
The mixed mark liquid chromatogram of 2 Fig. 5 c organic toxin of embodiment；Fig. 5 d wheat negative sample and negative mark-on sample compare
Figure.
The mixed mark liquid chromatogram of 3 Fig. 5 e organic toxin of embodiment；Fig. 5 f maize flour negative sample liquid chromatogram；Fig. 5 g is beautiful
Rice and flour feminine gender mark-on sample comparison diagram.
Appended drawing reference: 1. ultrasonic transducers；2, handle；3, pipette tips interface；4, imbibition key；5, drain key；6, pipette tips；
7, battery charge port.
Specific embodiment
The present invention provides organic toxin sample purification extraction detection method in a kind of cereal crops, and detecting step includes: (1)
Sample extraction: taking measuring samples Extraction solvent constant volume, mechanical shaking extraction, and collection supernatant is sample extracting solution；(2) sample is net
Change: pipette samples extracting solution quantify using liquid-transfering gun, make extracting solution flow through/flow through scavenging material, continue quantitative draw air, it is bulging
Bubble in dynamic sample extracting solution, makes scavenging material adsorbing contaminant in liquid transfer gun head；(3) it vibration extraction: is shaken using liquid-transfering gun ultrasound
Dynamic sample extracting solution is crushed bubble in scavenging material hole with sample extracting solution and mixes, makes scavenging material in gas-liquid interface
Further extracting impurities；(3) sample drain: purified sample is discharged, and is sent to machine testing on detection device；
Wherein, in step (2), sample purification step includes: 1. to take out pipette tips, is installed on liquid-transfering gun, and pipette tips are protruded into
Sample liquid level to be clean makes liquid-transfering gun generate negative-pressure liquid suction in pipette tips hereinafter, press liquid-transfering gun, sucks 1ml sample to be clean,
So that purification sample is flowed through/flow through purification sample, stops 5s；2. liquid-transfering gun pipette tips are removed liquid level, liquid-transfering gun is pressed again, makes to move
Negative-pressure liquid suction is generated in liquid rifle in pipette tips, 1ml air is sucked, agitates bubble in sample extracting solution, fill liquid with scavenging material
Divide mixing, repeat this step three times, sucks 4ml air liquid-transfering gun altogether and show that 5000ul ALL is fully loaded with；3. opening liquid-transfering gun ultrasound
4. pipette tips are directed at 2ml sample bottle, press liquid-transfering gun by energy converter, sample extracting solution and scavenging material in ultrasonic vibration liquid-transfering gun
Drain, machine analysis on collection and purification liquid.
Wherein, in step (1), extraction step includes: to weigh 5g cereal crops sample comminution, crosses 1mm sub-sieve, in
In 50mL centrifuge tube, disperses constant volume to 25mL with extracting solution (95wt% acetonitrile/water), be vortexed and mix；Ultrasonic extraction 30min, puts
Enter centrifuge (8000r/min is centrifuged 20min), takes supernatant as sample extracting solution.The cereal crops are preferably plant
Cereal crops, including corn, wheat and rice；Organic toxin includes zearalenone, patulin and vomitoxin.
Wherein, it in step (3), is analyzed by high performance liquid chromatography (HPLC-LC-20A), patulin detector bar
Part are as follows:
Mobile phase: A phase, acetonitrile；B phase, acetonitrile -0.5mM ammonium acetate (14:86, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 0.7mL/min；
Column temperature: 30 DEG C；
Sample volume: 20 μ L；
Fluorescence detection: excitation wavelength: 276nm；Launch wavelength: 440nm, photochemistry post-column derivation.
Wherein, it in step (3), is analyzed by high performance liquid chromatography (HPLC-LC-20A), vomitoxin detector bar
Part are as follows:
Mobile phase: methanol-water (20:80, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 1.0mL/min；
Column temperature: 35 DEG C；
Sample volume: 50 μ L；
Ultraviolet detection: excitation wavelength: 218nm.
Scavenging material includes but is not limited to: diatomite, magnesium sulfate, ketjenblack EC, sulfonic group n-vinyl pyrrolidone-
Divinylbenzene, carboxyl n-vinyl pyrrolidone-divinylbenzene, methylene piperazine ring n-vinyl pyrrolidone-diethyl
Alkenyl benzene, C18Modified silica-gel, C8Modified silica-gel, aminopropyl bonded silica gel, glycol-based bonded silica gel, florisil silica or aluminium oxide
At least one of.
The present invention also provides a kind of purifications suitable for above-mentioned detection method to extract liquid-transfering gun comprising gun body, gun body packet
Handle, ultrasonic transducer, imbibition key, drain key, pipette tips interface and battery pack are included, ultrasonic transducer pump tracheae connects with pipette tips
Mouth is connected, and ultrasonic transducer is electrically connected with imbibition key, drain key and battery pack respectively.
Detection implementation process is purified to the present invention below according to specific embodiment to be illustrated:
Embodiment 1
Zearalenone application case in maize flour
Sample extraction:
6g maize flour sample is weighed, crushes and crosses 1mm sub-sieve, be added in 50mL centrifuge tube, 0.6g sodium chloride is added
It is mixed with 15mL extracting solution (90% acetonitrile-aqueous solution).
Ultrasonic extraction 30min is put into centrifuge (8000r/min is centrifuged 20min), takes supernatant as sample extraction
Liquid.
Sample purification:
Step 1: taking out pipette tips, be installed on electrical pipette rifle, pipette tips are protruded into liquid liquid level to be clean hereinafter, clicking electricity
Dynamic liquid-transfering gun imbibition key, sucks 1ml liquid to be clean, stops 5s.
Step 2: liquid-transfering gun pipette tips being removed into liquid level, click electrical pipette rifle imbibition key again, sucking 1ml is empty
Gas is sufficiently mixed liquid with filler, repeats this step three times, sucks 4ml air altogether.
Step 3: opening liquid-transfering gun ultrasonic transducer, sample extracting solution and scavenging material in ultrasonic vibration liquid-transfering gun
3min,
Step 4: liquid-transfering gun shows 5000ul ALL, and pipette tips are directed at 2ml sample bottle, click liquid-transfering gun drain again
Key carries out drain, collection and purification liquid.
Upper machine analysis: sample purification liquid is sent to machine testing on HPLC detection device.
HPLC testing conditions:
Mobile phase: A phase, water；B phase, acetonitrile-methanol-water (46:8:46, v:v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 1.0mL/min；
Column temperature: 35 DEG C；
Sample volume: 50 μ L；
Fluorescence detection: excitation wavelength: 274nm；Launch wavelength: 440nm, photochemistry post-column derivation.
Experimental result:
Testing result: Fig. 5 a zearalenone liquid chromatogram.Wherein, zearalenone ZON concentration is 100ng/
mL。
The liquid chromatogram of Fig. 5 b maize flour sample extracting solution, wherein ZON concentration is 218.3ng/g.
1 maize flour sample minimum detectability of table, recovery of standard addition and repeatability (ZON concentration is 218.3ng/g)
* ZON is the abbreviation of zearalenone
Embodiment 2
Organic toxin application case in wheat
Sample extraction:
5g wheat samples are weighed, crushes and crosses 1mm sub-sieve, be added in 50mL centrifuge tube, it is fixed with extracting solution (acetonitrile)
Hold 25mL, is vortexed and mixes；
Ultrasonic extraction 30min is put into centrifuge (4000r/min is centrifuged 10min), takes supernatant as sample extraction
Liquid.
Sample purification:
Step 1: taking out pipette tips, be installed on electrical pipette rifle, pipette tips are protruded into liquid liquid level to be clean hereinafter, clicking electricity
Dynamic liquid-transfering gun imbibition key, sucks 1ml liquid to be clean, stops 5s.
Step 2: liquid-transfering gun pipette tips being removed into liquid level, click electrical pipette rifle imbibition key again, sucking 1ml is empty
Gas is sufficiently mixed liquid with filler, repeats this step three times, sucks 4ml air altogether.
Step 3: opening liquid-transfering gun ultrasonic transducer, sample extracting solution and scavenging material in ultrasonic vibration liquid-transfering gun
5min,
Step 4: liquid-transfering gun shows 5000ul ALL, and pipette tips are directed at 2ml sample bottle, click liquid-transfering gun drain again
Key carries out drain, collection and purification liquid.
Step 5: take 2mL purify extracting solution, nitrogen be blown to it is dry, with 1mL or 0.5mL initial liquid phase redissolve, cross micropore filter
Film, upper machine analysis.
HPLC testing conditions:
Mobile phase: A phase, acetonitrile；B phase, acetonitrile -0.5mM ammonium acetate (14:86, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 0.7mL/min；
Column temperature: 30 DEG C；
Sample volume: 20 μ L；
Fluorescence detection: excitation wavelength: 276nm；Launch wavelength: 440nm, photochemistry post-column derivation.
As a result it calculates:
Experimental result:
Fig. 5 c patulin liquid chromatogram, wherein patulin concentration is 1 μ g/mL.
Fig. 5 d wheat samples extracting solution liquid chromatogram, wherein patulin detection value is 0.09 μ g/g.
Fig. 5 e wheat samples extracting solution mark-on sample liquid chromatogram, wherein patulin spiked levels are 1.05 μ g/g.
(PAT concentration is for 2 wheat samples extracting solution patulin minimum detectability of table, recovery of standard addition and repeatability
218.3ng/g)
* PAT is the abbreviation of zearalenone
Embodiment 3
Vomitoxin application case in maize flour
Sample extraction:
5g maize flour sample is weighed, crushes and crosses 1mm sub-sieve, be added in 50mL centrifuge tube, use extracting solution
(84wt%, acetonitrile/water) constant volume is vortexed and mixes to 25mL；
Ultrasonic extraction 30min is put into centrifuge (8000r/min is centrifuged 20min), takes supernatant as sample extraction
Liquid.
Sample purification:
Step 1: taking out pipette tips, be installed on electrical pipette rifle, pipette tips are protruded into liquid liquid level to be clean hereinafter, clicking electricity
Dynamic liquid-transfering gun imbibition key, sucks 1ml liquid to be clean, stops 5s.
Step 2: liquid-transfering gun pipette tips being removed into liquid level, click electrical pipette rifle imbibition key again, sucking 1ml is empty
Gas is sufficiently mixed liquid with filler, repeats this step three times, sucks 4ml air altogether.
Step 3: opening liquid-transfering gun ultrasonic transducer, sample extracting solution and scavenging material in ultrasonic vibration liquid-transfering gun
3min,
Step 4: liquid-transfering gun shows 5000ul ALL, and pipette tips are directed at 2ml sample bottle, click liquid-transfering gun drain again
Key carries out drain, collection and purification liquid.
Step 5: take 2mL purify extracting solution, nitrogen be blown to it is dry, with 1mL or 0.5mL initial liquid phase redissolve, cross micropore filter
Film, upper machine analysis.
HPLC testing conditions:
Mobile phase: methanol-water (20:80, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 1.0mL/min；
Column temperature: 35 DEG C；
Sample volume: 50 μ L；
Ultraviolet detection: excitation wavelength: 218nm.
Experimental result:
Fig. 5 f vomitoxin liquid chromatogram, wherein DON concentration is 1000ng/mL.
Fig. 5 g maize flour sample extracting solution liquid chromatogram, DON concentration are 900ng/g；
Wherein, AFG1 and B1 spiked levels are 10ng/g；AFG2 and B2 spiked levels are 3ng/g.
2 maize flour sample minimum detectability of table, recovery of standard addition and repeatability (DON concentration is 900ng/g)
DON is vomitoxin abbreviation
All above-mentioned this intellectual properties of primarily implementation, there is no this new products of implementation of setting limitation other forms
And/or new method.Those skilled in the art will utilize this important information, above content modification, to realize similar execution feelings
Condition.But all modifications or transformation belong to the right of reservation based on new product of the present invention.
The above described is only a preferred embodiment of the present invention, being not that the invention has other forms of limitations, appoint
What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc.
Imitate embodiment.But without departing from the technical solutions of the present invention, according to the technical essence of the invention to above embodiments institute
Any simple modification, equivalent variations and the remodeling made, still fall within the protection scope of technical solution of the present invention.
Claims (7)
1. organic toxin sample purification extracts detection method in a kind of cereal crops, which is characterized in that detecting step includes: (1)
Sample extraction: taking measuring samples Extraction solvent constant volume, mechanical shaking extraction, and collection supernatant is sample extracting solution；(2) sample is net
Change: pipette samples extracting solution quantify using liquid-transfering gun, make extracting solution flow through/flow through scavenging material, continue quantitative draw air, it is bulging
Bubble in dynamic sample extracting solution, makes scavenging material adsorbing contaminant in liquid transfer gun head；(3) it vibration extraction: is shaken using liquid-transfering gun ultrasound
Dynamic sample extracting solution is crushed bubble in scavenging material hole with sample extracting solution and mixes, makes scavenging material in gas-liquid interface
Further extracting impurities；(3) sample drain: purified sample is discharged, and is sent to machine testing on detection device；
Wherein, in step (2), sample purification step includes: 1. to take out pipette tips, is installed on liquid-transfering gun, and pipette tips are protruded into net
Change sample liquid level hereinafter, pressing liquid-transfering gun, liquid-transfering gun is made to generate negative-pressure liquid suction in pipette tips, suck 1ml sample to be clean, makes net
Change sample and flow through/flow through purification sample, stops 5s；2. liquid-transfering gun pipette tips are removed liquid level, liquid-transfering gun is pressed again, makes liquid-transfering gun
Negative-pressure liquid suction is generated in interior pipette tips, 1ml air is sucked, agitates bubble in sample extracting solution, keeps liquid and scavenging material sufficiently mixed
It closes, repeats this step three times, suck 4ml air liquid-transfering gun altogether and show that 5000ul ALL is fully loaded；3. opening liquid-transfering gun ultrasonic transduction
4. pipette tips are directed at 2ml sample bottle, press liquid-transfering gun drain by device, sample extracting solution and scavenging material in ultrasonic vibration liquid-transfering gun,
Machine is analyzed on collection and purification liquid.
2. purification extraction detection method as described in claim 1, which is characterized in that in step (1), extraction step includes:
5g cereal crops sample comminution is weighed, 1mm sub-sieve is crossed, in 50mL centrifuge tube, is dispersed with extracting solution (95wt% acetonitrile/water)
Constant volume is vortexed and mixes to 20mL；Ultrasonic extraction 30min is put into centrifuge (8000r/min is centrifuged 20min), takes supernatant conduct
Sample extracting solution；The cereal crops are preferably plant cereal crops, including maize flour, wheat and maize flour；Organic toxin
Including zearalenone, patulin and vomitoxin.
3. purification extraction detection method as claimed in claim 1 or 2, which is characterized in that in step (3), pass through efficient liquid
Phase chromatography (HPLC) is analyzed, zearalenone testing conditions are as follows:
Mobile phase: A phase, water；B phase, acetonitrile-methanol-water (46:8:46, v:v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 1.0mL/min；
Column temperature: 35 DEG C；
Sample volume: 50 μ L；
Fluorescence detection: excitation wavelength: 274nm；Launch wavelength: 440nm, photochemistry post-column derivation.
4. purification extraction detection method as claimed in claim 1 or 2, which is characterized in that in step (3), pass through efficient liquid
Phase chromatography (HPLC) is analyzed, patulin testing conditions are as follows:
Mobile phase: A phase, acetonitrile；B phase, acetonitrile -0.5mM ammonium acetate (14:86, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 0.7mL/min；
Column temperature: 30 DEG C；
Sample volume: 20 μ L；
Fluorescence detection: excitation wavelength: 276nm；Launch wavelength: 440nm, photochemistry post-column derivation；
5. purification extraction detection method as claimed in claim 1 or 2, which is characterized in that in step (3), pass through efficient liquid
Phase chromatography (HPLC) is analyzed, vomitoxin testing conditions are as follows:
Mobile phase: methanol-water (20:80, v:v)；
Gradient elution condition: A, 68%；B, 32%；
Chromatographic column: C18 column (column length 250mm, column internal diameter 4.6mm, 5 μm of packing material size)；
Flow velocity: 1.0mL/min；
Column temperature: 35 DEG C；
Sample volume: 50 μ L；
Ultraviolet detection: excitation wavelength: 218nm.
6. as claimed any one in claims 1 to 3 purification extraction detection method, which is characterized in that scavenging material include but
It is not limited to: diatomite, magnesium sulfate, ketjenblack EC, sulfonic group n-vinyl pyrrolidone-divinylbenzene, carboxyl N- ethylene
Base pyrrolidones-divinylbenzene, methylene piperazine ring n-vinyl pyrrolidone-divinylbenzene, C18Modified silica-gel, C8Change
At least one of property silica gel, aminopropyl bonded silica gel, glycol-based bonded silica gel, florisil silica or aluminium oxide.
7. a kind of ultrasonic liquid-transfering gun suitable for the detection method as described in any one of claim 1-6, which is characterized in that it is wrapped
Gun body is included, gun body includes handle, ultrasonic transducer, imbibition key, drain key, pipette tips interface and battery pack, ultrasonic transducer
Ultrasonic probe is connected with pipette tips interface, and ultrasonic transducer is electrically connected with imbibition key, drain key and battery pack respectively, surpasses
Sonic transducer preferably uses 28khz60W ultrasonic vibrator.
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