CN110146619A - The high-flux detection method of four kinds of fungicide in fruits and vegetables - Google Patents
The high-flux detection method of four kinds of fungicide in fruits and vegetables Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The present invention relates to a kind of high-flux detection method of four kinds of fungicide in fruits and vegetables, step includes: sample preparation；Extraction obtains liquid to be clean；Multi-example synchronous purification；Debugging or optimization gaschromatographic mass spectrometry testing conditions；It is analyzed using gas chromatography tandem mass spectrometry, uses the content of Bravo, difenoconazole, 4 kinds of procymidone, triazolone fungicide in more reactive ion monitoring patterns and inner mark method ration analysis sample.Wherein Multi-example synchronous purification refers to using the DPX-RP pipette tips formula dispersed solid phase semi-automatic solid-phase extraction device of microextraction column combination multichannel to the synchronous synchronization air-breathing for carrying out extraction step of multiple samples, imbibition, exhaust, drain.Ensure the consistency of different sample purification processing operations, and helps to effectively shorten time of sample pre-treatments.
Description
Technical field
The invention belongs to the qualitative and quantitative detection fields of agricultural product residue, and in particular to a kind of to detect hundred in fruits and vegetables simultaneously
Bacterium is clear, triazolone, four kinds of procymidone, difenoconazole fungicide detection method.
Background technique
The pathogenic microorganisms such as fungi, virus to plant pest have 80,000 kinds or more, and global crops are due to phytopathy
Evil causes a large amount of underproduction, counts according to FAO (Food and Agriculture Organization of the United Nation) (FAO), and annual crops lose because of disease up to 10% -15%.Change
The use for learning fungicide is a kind of cost-effective method of controlling plant diseases.The disinfectant use in agriculture developed and used in recent years
Mainly there are triazole type, methoxy acrylic, amino acid derived species, amides and other class fungicide.These are agricultural to kill
Microbial inoculum activity is high, the mechanism of action is unique, environmentally friendly, is widely used in grain, fruits and vegetables, the gray mold of industrial crops, downy mildew
The prevention and treatment of the diseases such as disease.However, since chemistry security realizes the scarcity with knowledge, overdose and Use overrun and by safety
The problems such as interval is harvested, and to the protection of ambient soil and the attention of people's health, many countries are all formulated
Limit standard in agricultural product/food.Practice for Pesticide Residue in Agricultural Products testing result showed the fungicide drugs such as Bravo in recent years
It is one of higher pesticide of recall rate, residue problem is increasingly subject to the concern of people.Exploitation is high-throughput and scene is fast in real time
Fast detection technique, for ensureing that agricultural product security is of great significance.
At present for the analysis of multicomponent pesticide residue, gas-chromatography, liquid chromatogram or its mass spectrometry skill are mainly used
Art carries out separation detection.In recent years gas chromatography tandem mass spectrometry method (GC-MS/MS) has obtained biggish popularization, high sensitivity,
High specificity.The extraction of sample mostly uses the methods of oscillation homogeneous, supercritical extract, accelerated solvent extraction.The purification of sample is adopted
Sample purification is carried out with the methods of gel permeation chromatography, Solid Phase Extraction, QuEChERS (referring to publication number CN105203661A).
DPX pipette tips formula dispersed solid phase microextraction column is a kind of novel solid phase extraction techniques, passes through pipette tips formula device, dispersion
Solid Phase Extraction speed has been significantly increased in type resin extraction technology, while effectively reducing the dosage of solvent.DPX pipette tips formula point
The absorption resin for dissipating filling dispersion inside solid phase microextraction column, under the conditions of appropriate solvent, specific adsorption purpose compound, warp
Non-specific adsorption impurity can be removed by cleaning buffer washing, then can make purpose sample from corresponding resin with elution buffer elution
The purifying and concentration of purpose sample are completed in separation.Compared with dispersive solid-phase extraction method, good purification, impurity interference is less.DPX
The advantages of pipette tips formula dispersed solid phase microextraction column combines solid phase extraction and dispersive solid-phase extraction method, in sample impurity it is net
Change thorough.
When detecting multiple samples, using the pretreatment mode of the non-extraction column of existing dispersed solid phase, there are time length, low efficiency
The problem of, multiple samples can not synchronization process, and exist and completely the same can not lead to the risk of error because of processing operation.
Summary of the invention
The technical problem to be solved by the present invention is to provide in a kind of fruits and vegetables four kinds of fungicide for the above-mentioned prior art
High-flux detection method, for multiple samples, can the multiple samples of simultaneously operating pre-treatment, be based especially on DPX pipette tips formula point
Dissipate the synchronization sample treatment of solid phase microextraction column purifying step, it is ensured that the pre-treatment of multiple samples is completely the same, improves processing effect
Rate and accuracy in detection.
The present invention solves the above problems used technical solution are as follows: the high-throughput detection side of four kinds of fungicide in fruits and vegetables
Method, step include
(1) prepared by sample
The edible portion of fruits and vegetables sample is taken to shred, smash to pieces, homogeneous mixes, sealing, as sample, stored frozen；
(2) it extracts
Sample is weighed in centrifuge tube, chlopyrifos-D is accurately added10Acetonitrile, mechanical shaking extraction, centrifuging and taking is added in inner mark solution
Saturated sodium chloride solution and deionized water is added, as liquid to be clean in supernatant；
(3) Multi-example synchronous purification
According to step (1) and (2) if preparation dry-eye disease liquid to be clean, using Multi-channel solid phase extraction device to it is multiple to
Scavenging solution synchronizes purification, and purifying step includes:
(3.1) balance activation: prepare the DPX-RP pipette tips formula dispersed solid phase microextraction column equal with sample size, from DPX-
Acetonitrile is drawn in the pipette tips formula dispersed solid phase microextraction column bottom end RP, and then all discharges, are repeated 1 times or repeatedly；
(3.2) sampling combines: several extraction columns continue air-breathing shape from the sample to be clean in bottom end sucking step (2)
At turbulent flow, liquid is all discharged after combining for absorption；
(3.3) wash: several DPX-RP pipette tips formula dispersed solid phase microextraction columns suck deionized water from bottom end, then by liquid
Body is all discharged, and is repeated 1 times or repeatedly；
(3.4) elute: several DPX-RP pipette tips formula dispersed solid phase microextraction columns are mixed from bottom end sucking ethyl acetate-hexane
Close solution, liquid all exported into sample bottle, static a moment, allow liquid layered, another supernatant liquid of drawing into interpolation pipe,
It is accurate that Heptachlor epoxide inner mark solution is added；
(4) debug or optimize gaschromatographic mass spectrometry testing conditions；
(5) it is analyzed using gas chromatography tandem mass spectrometry, is analyzed using more reactive ion monitoring patterns and inner mark method ration
The content of 4 kinds of Bravo, difenoconazole, procymidone, triazolone fungicide in sample.
The Multi-example synchronous purification that above-mentioned steps (3) are mentioned refer to using Multi-channel solid phase extraction device to multiple samples into
The operation of the synchronization imbibition of row extraction step or air-breathing or exhaust, drain.
Mass Spectrometer Method condition optimizing is by four kinds of target detection fungicide, chlopyrifos-D in step (4)10Internal standard, ring
Oxygen heptachlor internal standard carries out full scan, obtains retention time and parent ion, then by Product ion scans, optimization daughter ion and touches
Hit energy.
Step (5) quantitative analysis uses calibration curve method, in order to eliminate background interference, prepares standard using bare substrate liquid
Curve makes calibration curve with the standard solution that the sample purification liquid of blank prepares four fungicide, carries out quantitative analysis and meter
It calculates.
Compared with the prior art, the advantages of the present invention are as follows:
(1) sample pre-treatments are filled by the semi-automatic Solid Phase Extraction of DPX pipette tips formula dispersed solid phase microextraction column combination multichannel
It sets and synchronous purification processing is made to multiple samples, it is ensured that the consistency of purified treatment operation, it is time saving, laborsaving, improve pre-treatment
Operating efficiency.The triple quadrupole rods tandem mass spectrometries of gas-chromatography-measure Bravo in fruits and vegetables, difenoconazole, procymidone, three simultaneously
The content of 4 kinds of fungicide of oxazolone.
(2) bare substrate liquid preparing standard solution and more reaction detections (MRM) mode are used, matrix effect is reduced, mentions
The high accuracy and sensitivity of analysis, test result are more accurate.
(3) water content of fruits and vegetables sample is higher, the low polar solvents such as acetonitrile can be selected as extraction of about 75%
Agent, both can be improved the immersional wetting of extractant, also can be reduced the water content in extracting solution, be conducive to subsequent purification.
(4) after extraction conditions of the sample by optimization extract, select DPX-RP pipette tips formula dispersed solid phase microextraction column to mesh
Mark object is purified, and loose resin is filled in liquid transfer gun head by DPX pipette tips formula dispersed solid phase microextraction column, passes through pipette tips formula
Solid Phase Extraction speed has been significantly increased in device, dispersion type resin abstraction technique, while effectively reducing the dosage of solvent.It is real
Test the result shows that, by purification after, target compound is retained, and the impurity in sample has obtained preferable removal.DPX rifle
Hair style dispersed solid phase microextraction column biggest advantage is that step is simple, effectively reduces the usage amount of organic solvent, pipette tips end
Sieve apparatus can filter out particulate matter, thus eliminate centrifugation etc. removal particulate matter operation.
(5) time for utilizing the semi-automatic solid-phase extraction device of multichannel, while handling 1-24 samples is 10-20 points
Clock, high-throughput, purification efficiency is high, and each extraction column synchronizes purification run to sample, farthest realizes that all samples are net
Change uniformity for the treatment of.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of solid-phase extraction device of the embodiment of the present invention, is not drawn into trachea in figure；
Fig. 2 is the structural schematic diagram of sample of embodiment of the present invention mechanism；
Fig. 3 is the structural schematic diagram of pipetting device of the embodiment of the present invention；
Fig. 4 is the partial structure diagram of pipetting device of the embodiment of the present invention；
Fig. 5 is the structural schematic diagram of the porous supporting plate of the embodiment of the present invention；
Fig. 6 is the structural schematic diagram of seal cover board of the embodiment of the present invention；
Fig. 7 is the structural schematic diagram of test solution of embodiment of the present invention box；
Fig. 8 is the structural schematic diagram of extraction column in the embodiment of the present invention.
Specific embodiment
The present invention will be described in further detail below with reference to the embodiments of the drawings.
Embodiment 1
Based on Bravo, difenoconazole, procymidone, 4 kinds of triazolone sterilizations in GC-MS/MS method detection veterinary antibiotics
The detection method of agent content, step include
One, sample pre-treatments
(1.1) sampling and sample preparation
Tomato, apple, Xiaoqinling Nature Reserve pick up from local supermarket, the market of farm produce.It takes edible portion to shred, is smash with high-speed organization
Broken machine, homogeneous mix, and sealing is spare as sample, are placed in -20 DEG C of preservations.
(1.2) it extracts
10g sample is weighed, is accurate to 0.01g in 50mL centrifuge tube, it is accurate that 40 μ L chlopyrifos-D are added10Inner mark solution
10mL acetonitrile is added in (Isotopic Internal Standard object), is placed in oscillation vortice shaking 15min, is centrifuged 10min in 4000r/min.Accurately
2.5mL supernatant is drawn in test tube, 2.5mL saturated sodium chloride solution and 6mL deionized water is added, as liquid to be clean.
(1.3) it purifies
The purification of sample passes through the semi-automatic Solid Phase Extraction dress of DPX-RP pipette tips formula dispersed solid phase microextraction column combination multichannel
Set draw solution or sample.Specific step is as follows:
According to step (1.1) and (1.2) if the liquid to be clean of preparation dry-eye disease, using Multi-channel solid phase extraction device to more
A liquid to be clean synchronizes purification, and purifying step includes:
(1) balance activation: connection 5mL DPX-RP pipette tips formula dispersed solid phase microextraction column and the semi-automatic solid phase extraction of multichannel
Device is taken, multiple extraction columns suck 1mL acetonitrile from extraction column bottom end, are inhaled after making a call to 3 times repeatedly, stand 30 seconds, and liquid is all arranged
Out；(2) sampling combines: (about with the sample to be clean in DPX-RP pipette tips formula dispersed solid phase microextraction column aspiration step (1.2)
12mL), divide 4 absorptions, draw the liquid to be clean of 3mL or so every time, be lifted away from liquid level, make the micro- extraction of DPX-RP pipette tips formula dispersed solid phase
It takes column more than liquid liquid level to be clean, then proceedes to air-breathing, form turbulent flow in extraction column, 30 seconds are stood, by liquid in conjunction with after
Discharge, then imbibition, air-breathing, standing, drain again repeat this operation totally 4 times；(3) wash: extraction column sucks 1mL from bottom end
Deionized water inhales after making a call to 3 times repeatedly, stands 30 seconds, and liquid is all discharged；(4) it elutes: sucking 700 μ L from extraction column bottom end
Ethyl acetate-hexane (1:1, V:V) mixed solution, repeatedly inhale make a call to 3 times after, stand 30 seconds, then by liquid all discharges
Into sample bottle, in static a moment, liquid layered is allowed, draw supernatant liquid (organic phase) 300 μ L and, into interpolation pipe, be accurately added 12
μ L Heptachlor epoxide inner mark solution is analyzed with gas chromatography tandem mass spectrometry (GC-MS/MS).
Using the semi-automatic solid-phase extraction device of multichannel, 24 samples can be handled simultaneously, required time is 10-20 minutes,
High throughput, purification efficiency is higher, and imbibition, air-breathing, the drain of 24 samples operate fully synchronized flux.
Two, gas chromatography tandem mass spectrometry (GC-MS/MS) operating condition
(1) chromatographic condition
Agilent HP-5MS Ultra Inert chromatographic column (30m × 0.25mm × 0.25 μm)；Temperature program: 60 DEG C of holdings
1min rises to 170 DEG C with 40 DEG C/min, then rises to 310 DEG C of holding 3min with 10 DEG C/min；Sample volume: 1 μ L；Carrier gas: helium,
Purity >=99.999%, flow velocity: 1.0mL/min；Injector temperature: 250 DEG C；Input mode: Splitless injecting samples.
(2) Mass Spectrometry Conditions
Using electron impact ion source (EI)；Electron energy: 70eV；Ion source temperature: 300 DEG C；The solvent delay time:
4.08min；Scanning mode: more reactive ions monitor (MRM) mode.Retention time, the fragment ion information, impact energy of each component
The table 1 referring to summary of the invention is measured, wherein " * " is quantitative.Error when Heptachlor epoxide is analyzed for detecting instrument, and according to condition
Establish multiple-reaction monitoring method.
Retention time, the fragment ion information, collision energy of 1 fungicide of table
Three, the range of linearity of detection method, detection limit and quantitative limit
Using calibration curve method to each fungicide carry out quantitative analysis, using bare substrate liquid prepare standard curve, i.e., with
The scavenging solution of blank prepares corresponding standard solution and makes calibration curve, weighs 7 parts of blank samples, carries out according to step (1-3)
The bare substrate matching criteria working solution that mass concentration is 10,20,50,100,250,500,800 μ g/L is prepared in pre-treatment respectively
It is measured, with the quota ion peak area of each fungicide and corresponding internal standard chlopyrifos-D10Peak area ratio is ordinate Y,
Linear regression is done to mass concentration X, unit μ g/L, obtains the linear equation and related coefficient of each component pesticide.
It is that 3 (S/N=3) determine detection limit (LOD) with signal-to-noise ratio, with signal-to-noise ratio for 10 (S/ using the method for mark-on reclaims
N=10 quantitative limit (LOQ)) is determined, as a result referring to table 2, as seen from table, and in the concentration range of 10-800 μ g/L, 4 kinds of sterilizations
The linearly dependent coefficient of agent is all larger than 0.999, shows good linear relationship, and the detection of 4 kinds of fungicide is limited to 0.3-2.3 μ
G/kg is quantitatively limited to 1.2-6.7 μ g/kg, shows higher sensitivity.
2 fungicide linear dimensions of table, detection limit and quantitative limit
Four, the rate of recovery and precision discussion
Under experiment condition after optimization, mark-on reclaims test is done in tomato, apple and Xiaoqinling Nature Reserve blank sample respectively,
It is mixed that addition wherein is equivalent to fungicide of the sample containing 50 basic, normal, high 3 concentration levels of μ g/kg, 250 μ g/kg and 500 μ g/kg
Mark, each mark-on level carry out 6 parallel analysis respectively, calculate average recovery rate and relative standard deviation, the results are shown in Table 3.3
Under a pitch-based sphere, the average recovery rate of 4 kinds of fungicide is 87.05% -115.81%, and relative standard deviation (RSD) is
0.29% -8.78%, show that the accuracy of the detection method of the application and precision can reach satisfactory result, testing result
With high reliability.
Each fungicide recovery of standard addition of table 3 and standard deviation
Embodiment two
DPX-RP pipette tips formula dispersed solid phase microextraction column combination multichannel semi-automatic solid-phase extraction device (this paper other parts
Also become Multi-channel solid phase extraction column), it is made of pipetting device 1 and sample mechanism 2, the similar note of the aspiration column 102 of pipetting device 1
24 pipette tips formula dispersed solid phase microextraction columns of the principle of emitter, multiple aspiration columns and sample mechanism realize that extraction column one is a pair of
It answers, when multiple aspiration column synchronization actions, it can be achieved that flux imbibition, air-breathing, drain, the exhaust of 24 extraction columns, it is ensured that extraction
The consistency of operation.
Pipetting device includes 101,24 consistent aspiration columns 102 of specification of liquid relief orifice plate, and any aspiration column 102 includes outer cylinder
1021 and piston 1022, the pull end of piston 1022 be provided with push jack 1024, be similar to syringe principle, piston 1022 is removable
Ground is arranged in outer cylinder 1021, and 1021 front end of outer cylinder is provided with gas nozzle 1023, the gas nozzle air-breathing when piston is taken out outside outer tube inner wall.
Outer cylinder 1021, which is fixed at level and empties, to be arranged on liquid relief orifice plate 101, in order to realize 24 aspiration columns 102
Synchronism, it is two groups that 24 aspiration columns 102, which are divided to, and each group of 12 aspiration columns 102 linearly arrange, and two groups of aspiration columns are mutually flat
Row.Meanwhile synchronous piece 103 has " I " fonts section, because having two bar shaped elongated slots, 12 push jacks linearly arranged
In 1024 corresponding card one of them bar shaped elongated slots, other 12 push jacks 1024 linearly arranged are stuck in another bar shaped elongated slot
It is interior.
In addition, operating convenient for the push-and-pull to synchronous piece 103, push-pull configuration, setting one are further configured in pipetting device
The top of column, column is higher than liquid relief orifice plate 102, and intermeshing rack gear 104, gear shaft 105, gear is arranged in column top
Axis 105 is configured with the synchronous piece 103 that the rack gear 104 that rotating handle 106 is vertically arranged is connected to lower section, 105 sliding tooth of gear shaft
Item 105 lifting with pull up or under push away synchronous piece 103.
Sample mechanism 2 includes sample mount 201, porous supporting plate 202, sealing clamp 203, and porous 202 level of supporting plate is emptied
It is arranged in sample mount 201,24 pipette tips formula dispersed solid phase microextraction columns 3 are suspended on porous supporting plate 202, and extraction column point
For parallel two rows, a row 12.It is provided with 24 on sealing clamp 203 and connects stomata 2031, there are two the porous settings of supporting plate 202
Positioning column 2021, correspondingly there are two location holes 2032 for setting on sealing clamp 203, are worn by positioning column to location hole alignment
Hole makes sealing clamp 203 and porous supporting plate 202, and accurately lid closes, and Gai Hehou, connects stomata 2031 and the micro- extraction of DPX pipette tips formula dispersed solid phase
It takes the ventilation corking at 3 top of column to correspond connection and forms gas circuit.In order to improve leakproofness, location hole is passed through in positioning column 2021
2032, being exposed on the positioning column 2021 of location hole 2032 makes porous supporting plate 202 and sealing clamp 203 by the way that lock tube 2022 is arranged
Close apposition.
The gas nozzle 1023 of 24 aspiration columns (is not drawn into figure by 24 piece-root grafting tracheaes, but is not difficult to manage according to recording herein
The solution trachea is hollow plastic tube) other ends that connect stomata 2031 with 24 correspond and connect, in synchronous piece 103
24 pipette tips formula dispersed solid phase microextraction columns 3 synchronous air-breathing when pull piston 1022 simultaneously, and inspiratory capacity is consistent.
Come again, sample mechanism 2 further includes a flute profile test solution box 4, and test solution box 4 is placed in pipette tips formula dispersed solid phase microextraction column
3 lower sections, for accepting the test solution of single or multiple pipette tips formula dispersed solid phase microextraction column outflows.
In order to match the pipette tips formula dispersed solid phase microextraction column of different length, the bearing height of more sky supporting plates should be set as
Adjustable: porous 202 both ends of supporting plate are provided with sliding sleeve 2023, and two sliding sleeves 2023 are through 201 or so two root posts of sample mount
On 2011, two sets can be slided up and down along column 2011 to adjust the height of porous supporting plate 202.
The aspiration column of large capacity is used in the present embodiment, such as 20mL can suck always gas in a longer period of time,
The time for forming turbulent flow is long, and mixing is more abundant, and the effect of extracting and purifying is more preferable.
The semi-automatic solid-phase extraction device of DPX-RP pipette tips formula dispersed solid phase microextraction column combination multichannel of the present embodiment can
Realize the column operation of the synchronous flux imbibition of the sample in 24 channels, air-breathing, drain, exhaust, it is ensured that each sample is handled consistent
Property, certainly, according to actual needs, 24 channels can also be designed as to 12 channels or other specifications.
In addition to the implementation, all to use equivalent transformation or equivalent replacement the invention also includes there is an other embodiments
The technical solution that mode is formed should all be fallen within the scope of the hereto appended claims.
Claims (10)
1. the high-flux detection method of four kinds of fungicide in a kind of fruits and vegetables, it is characterised in that: step includes
(1) prepared by sample
The edible portion of fruits and vegetables sample is taken to shred, smash to pieces, homogeneous mixes, sealing, as sample, stored frozen；
(2) it extracts
Sample is weighed in centrifuge tube, chlopyrifos-D is accurately added10Acetonitrile, mechanical shaking extraction, centrifuging and taking supernatant is added in inner mark solution
Saturated sodium chloride solution and deionized water is added, as liquid to be clean in liquid；
(3) Multi-example synchronous purification
According to step (1) and (2) if the liquid to be clean of preparation dry-eye disease, using Multi-channel solid phase extraction device to multiple to be clean
Liquid synchronizes purification, and purifying step includes:
(3.1) balance activation: prepare the DPX-RP pipette tips formula dispersed solid phase microextraction column equal with sample size, from DPX-RP rifle
Acetonitrile is drawn in hair style dispersed solid phase microextraction column bottom end, and then all discharges, are repeated 1 times or repeatedly；
(3.2) sampling combines: several extraction columns continue air-breathing and form rapids from the sample to be clean in bottom end sucking step (2)
Liquid is all discharged after combining for stream, absorption；
(3.3) wash: several DPX-RP pipette tips formula dispersed solid phase microextraction columns suck deionized water from bottom end, then that liquid is complete
Portion's discharge, is repeated 1 times or repeatedly；
(3.4) elute: several DPX-RP pipette tips formula dispersed solid phase microextraction columns are molten from bottom end sucking ethyl acetate-hexane mixing
Liquid all exports liquid into sample bottle, static a moment, allows liquid layered, and another supernatant liquid of drawing is into interpolation pipe, accurately
Heptachlor epoxide inner mark solution is added；
(4) debug or optimize gaschromatographic mass spectrometry testing conditions；
(5) it is analyzed using gas chromatography tandem mass spectrometry, analyzes sample using more reactive ion monitoring patterns and inner mark method ration
In Bravo, difenoconazole, 4 kinds of procymidone, triazolone fungicide content.
2. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 1, it is characterised in that: Multi-example is same
Step purification refer to using Multi-channel solid phase extraction device to multiple samples carry out extraction step synchronization imbibition or air-breathing or exhaust,
The operation of drain, Multi-channel solid phase extraction device include pipetting device (1) and sample mechanism (2), and the pipetting device includes moving
Fluid apertures plate (101), several aspiration columns (102) and synchronous piece (103),
The aspiration column (102) includes outer cylinder (1021) and piston (1022), and the piston (1022) is movably disposed at institute
It states in outer cylinder (1021), outer cylinder (1021) front end is provided with gas nozzle (1023), the gas nozzle when piston is taken out outside outer tube inner wall
Air-breathing；The outer cylinder (1021) is fixed in the mounting hole of liquid relief orifice plate (101)；The piston (1022) of several aspiration columns point
Not by the synchronous piece (103) pari passu pull or propulsion；
The sample mechanism (2) includes sample mount (201), porous supporting plate (202), sealing clamp (203), the porous supporting plate
(202) level, which is emptied, is arranged on the sample mount (201), and several DPX-RP pipette tips formula dispersed solid phase microextraction columns (3) are outstanding
Hang on the porous supporting plate (202), the sealing clamp (203) if on be provided with stem grafting stomata (2031), in the sealing
When pressing plate (203) and porous supporting plate (202) lid close, if one end of stem grafting stomata (2031) and the dispersion of several DPX-RP pipette tips formulas are solid
Phase extraction column (3), which corresponds, to be connected；
Described porous supporting plate (202) both ends are provided with sliding sleeve (2023), and the sample mount (201) has two root posts
(2011), two sliding sleeves (2023), which respectively correspond, covers on two root posts (2011), and can be along column (2011) up and down
It slides to adjust the height of porous supporting plate (202)；
The gas nozzle (1023) of several aspiration columns is connected if being corresponded by trachea and the other end of stem grafting stomata (2031),
Several pipette tips formula dispersed solid phase microextraction columns (3) synchronize air-breathing when the synchronous piece (103) while pull piston (1022)
Or outlet.
3. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 1, it is characterised in that: step (5)
Quantitative analysis uses calibration curve method, prepares standard curve using bare substrate liquid, prepares four with the sample purification liquid of blank
The standard solution of fungicide makes calibration curve, weighs 7 parts of blank samples, carries out pre-treatment according to step (1-3), prepares respectively
Mass concentration is that the bare substrate matching criteria working solution of 10,20,50,100,250,500,800 μ g/L is measured, respectively to kill
The quota ion peak area of microbial inoculum and corresponding internal standard chlopyrifos-D10Peak area ratio is ordinate Y, makees line to mass concentration X
Property return, obtain the linear equation and related coefficient of each component pesticide；
It is that S/N=3 determines detection limit with signal-to-noise ratio using the method for mark-on reclaims, is that S/N=10 determines quantitative with signal-to-noise ratio
Limit, as a result referring to following table, as seen from table, in the concentration range of 10-800 μ g/L, the linearly dependent coefficient of 4 kinds of fungicide is equal
Greater than 0.999, good linear relationship is showed, the detection of 4 kinds of fungicide is limited to 0.3-2.3 μ g/kg, is quantitatively limited to 1.2-
6.7 μ g/kg, show higher sensitivity；
4. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 1, it is characterised in that: step (2)
Concrete operations: weigh 10.00g sample, it is accurate that 40 μ L chlopyrifos-D are added in 50mL centrifuge tube10Inner mark solution is added
10mL acetonitrile is placed in oscillation vortice shaking 15min, is centrifuged 10min in 4000r/min.Accurate 2.5mL supernatant of drawing is in examination
2.5mL saturated sodium chloride solution and 6mL deionized water is added, as liquid to be clean in Guan Zhong.
5. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 1, it is characterised in that: step (3)
Purifying step based on Solid Phase Extraction is
(1) balance activation: connection 5mL DPX-RP pipette tips formula dispersed solid phase microextraction column and the semi-automatic Solid Phase Extraction dress of multichannel
It sets, multiple extraction columns suck 1mL acetonitrile from extraction column bottom end, are inhaled after making a call to 3 times repeatedly, stand 30 seconds, and liquid is all discharged；
(2) sampling combines: with the sample to be clean in DPX-RP pipette tips formula dispersed solid phase microextraction column aspiration step (1.2)
(about 12mL), point 4 absorptions, draws the liquid to be clean of 2.5-3.5mL or so every time, is lifted away from liquid level, make DPX-RP pipette tips formula point
Solid phase microextraction column is dissipated more than liquid liquid level to be clean, then proceedes to air-breathing, forms turbulent flow in extraction column, stands 30 seconds, knot
Liquid is discharged after conjunction, then imbibition, air-breathing, standing, drain again, repeats this operation totally 4 times；
(3) wash: extraction column sucks 1mL deionized water from bottom end, is inhaled after making a call to 3 times repeatedly, stands 30 seconds, and liquid is all arranged
Out；
(4) it elutes: sucking ethyl acetate-hexane (1:1, V:V) mixed solution of 700 μ L from extraction column bottom end, inhale repeatedly and make a call to 3
After secondary, 30 seconds are stood, is then all expelled to liquid in sample bottle, static a moment, allows liquid layered, draws supernatant liquid
(organic phase) 300 μ L is accurate that 12 μ L Heptachlor epoxide inner mark solutions are added into interpolation pipe, with gas chromatography tandem mass spectrometry (GC-
MS/MS it) analyzes.
6. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 2, it is characterised in that: the piston
Pull end be provided with push jack (1024), the synchronous piece (103), which has, blocks the card slots of the push jack (1024).
7. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 6, it is characterised in that: described several
Aspiration column (102) linearly arranges, and the card slot is bar shaped elongated slot, and several push jacks (1024) linearly arranged are stuck in institute
It states in bar shaped elongated slot.
8. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 2, it is characterised in that: the liquid relief
Mechanism (1) further includes the push-pull configuration for acting on synchronous piece (103), the push-pull configuration include intermeshing rack gear (104),
Gear shaft (105), rack gear (104) connect synchronous piece (103), and gear shaft (105) drive rack (105) pushes and pulls synchronous piece (103),
The gear shaft (105) is configured with rotating handle (106).
9. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 2, it is characterised in that: described porous
Supporting plate (202) is provided with positioning column (2021), is provided with location hole (2032), the positioning column on the sealing clamp (203)
(2021) corresponding with location hole (2032) position；The positioning column (2021) passes through location hole (2032), is exposed to location hole
(2032) porous supporting plate (202) and sealing clamp (203) pairing are locked by the way that lock tube (2022) are arranged on positioning column (2021)
It is fixed.
10. the high-flux detection method of four kinds of fungicide in fruits and vegetables according to claim 2, it is characterised in that: if described
Dry DPX-RP pipette tips formula dispersed solid phase microextraction column (3) bottom end is air entry, and top is provided with ventable corking (301), institute
It states corking (301) and heads into and connect stomata (2031) with the sealing clamp (203).
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| CN111855366A (en) * | 2020-06-29 | 2020-10-30 | 中国水产科学研究院 | Automatic filtering type purification device for sample pretreatment |
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