CN107389829A - Method and device that is a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos - Google Patents
Method and device that is a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos Download PDFInfo
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- 210000002700 urine Anatomy 0.000 title claims abstract description 52
- 239000002207 metabolite Substances 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 37
- SBPBAQFWLVIOKP-UHFFFAOYSA-N chlorpyrifos Chemical compound CCOP(=S)(OCC)OC1=NC(Cl)=C(Cl)C=C1Cl SBPBAQFWLVIOKP-UHFFFAOYSA-N 0.000 title claims abstract description 27
- 239000005944 Chlorpyrifos Substances 0.000 title claims abstract description 25
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000001704 evaporation Methods 0.000 claims abstract description 19
- 238000010257 thawing Methods 0.000 claims abstract description 19
- 238000012360 testing method Methods 0.000 claims abstract description 16
- 238000000746 purification Methods 0.000 claims abstract description 15
- UCQFCFPECQILOL-UHFFFAOYSA-N diethyl hydrogen phosphate Chemical compound CCOP(O)(=O)OCC UCQFCFPECQILOL-UHFFFAOYSA-N 0.000 claims abstract description 14
- PKUWKAXTAVNIJR-UHFFFAOYSA-M O,O-diethyl thiophosphate Chemical compound CCOP([O-])(=S)OCC PKUWKAXTAVNIJR-UHFFFAOYSA-M 0.000 claims abstract description 12
- 238000000605 extraction Methods 0.000 claims abstract description 8
- 238000001819 mass spectrum Methods 0.000 claims abstract description 7
- 150000001875 compounds Chemical class 0.000 claims abstract description 6
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 87
- 150000002500 ions Chemical class 0.000 claims description 41
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 36
- 238000003860 storage Methods 0.000 claims description 34
- 239000007789 gas Substances 0.000 claims description 31
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 22
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 18
- 229910052757 nitrogen Inorganic materials 0.000 claims description 12
- 239000007921 spray Substances 0.000 claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000004949 mass spectrometry Methods 0.000 claims description 7
- 238000004704 ultra performance liquid chromatography Methods 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 6
- 238000005070 sampling Methods 0.000 claims description 6
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 5
- 230000004060 metabolic process Effects 0.000 claims description 5
- 238000005507 spraying Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
- 239000003480 eluent Substances 0.000 claims description 4
- 238000011068 loading method Methods 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 238000011049 filling Methods 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
- 238000004811 liquid chromatography Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 21
- 230000008901 benefit Effects 0.000 abstract description 4
- 238000004458 analytical method Methods 0.000 abstract description 3
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 abstract 2
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 abstract 1
- 239000012071 phase Substances 0.000 description 24
- 238000011084 recovery Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- QPCDCPDFJACHGM-UHFFFAOYSA-N N,N-bis{2-[bis(carboxymethyl)amino]ethyl}glycine Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC(O)=O QPCDCPDFJACHGM-UHFFFAOYSA-N 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 3
- 210000003169 central nervous system Anatomy 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 238000000589 high-performance liquid chromatography-mass spectrometry Methods 0.000 description 3
- 210000002381 plasma Anatomy 0.000 description 3
- 238000004445 quantitative analysis Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 238000003912 environmental pollution Methods 0.000 description 2
- 238000004817 gas chromatography Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 238000012113 quantitative test Methods 0.000 description 2
- -1 thio Diethyl phosphate Chemical compound 0.000 description 2
- NRKYWOKHZRQRJR-UHFFFAOYSA-N 2,2,2-trifluoroacetamide Chemical compound NC(=O)C(F)(F)F NRKYWOKHZRQRJR-UHFFFAOYSA-N 0.000 description 1
- GMWHTUNMFTUKHH-NDUABGMUSA-N 2-[2-(2-methoxyethoxy)ethoxy]ethyl (e)-2-cyano-3-(6-piperidin-1-ylnaphthalen-2-yl)prop-2-enoate;hydrochloride Chemical compound Cl.C1=CC2=CC(/C=C(C(=O)OCCOCCOCCOC)\C#N)=CC=C2C=C1N1CCCCC1 GMWHTUNMFTUKHH-NDUABGMUSA-N 0.000 description 1
- 102000012440 Acetylcholinesterase Human genes 0.000 description 1
- 108010022752 Acetylcholinesterase Proteins 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 229940022698 acetylcholinesterase Drugs 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 125000000217 alkyl group Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003139 biocide Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 231100000085 chronic toxic effect Toxicity 0.000 description 1
- 208000010877 cognitive disease Diseases 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 231100000571 death by poisoning Toxicity 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001212 derivatisation Methods 0.000 description 1
- 125000004177 diethyl group Chemical group [H]C([H])([H])C([H])([H])* 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000001211 electron capture detection Methods 0.000 description 1
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- 150000002148 esters Chemical class 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- BHEPBYXIRTUNPN-UHFFFAOYSA-N hydridophosphorus(.) (triplet) Chemical compound [PH] BHEPBYXIRTUNPN-UHFFFAOYSA-N 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000002917 insecticide Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 238000002552 multiple reaction monitoring Methods 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 150000002825 nitriles Chemical class 0.000 description 1
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 1
- 239000002957 persistent organic pollutant Substances 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000004451 qualitative analysis Methods 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 238000000825 ultraviolet detection Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000012224 working solution Substances 0.000 description 1
Classifications
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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
-
- 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
- G01N30/06—Preparation
-
- 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
- G01N30/06—Preparation
- G01N30/08—Preparation using an enricher
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- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of while detect the method and device of 3 kinds of metabolites of urine Chlorpyrifos.It the described method comprises the following steps：(A) target compound selects：The pyridine alcohol of 3,5,6 trichlorine 2, diethyl phosphate, diethyl thiophosphate；(B) sample digests；(C) sample extraction；(D) sample purification and concentration；(E) Instrumental Analysis：Described device mainly includes：Main body and chromatogram-and from tandem mass spectrum detector, include supply unit, thawing apparatus, centrifugal device one, water bath device, vortex mixed device, centrifugal device two, purifier, rotary evaporating device, PLC in the main body.The device of the present invention has the advantages of succinct quick, realizes intelligentized control method, and controllability is strong, and the degree of accuracy is high.The method of the present invention substantially increases the validity and reliability of detection, saves detection time and testing cost.
Description
Technical field
The present invention relates to Pesticides Testing technical field, is specifically related to a kind of while detects the 3 kinds of metabolism of urine Chlorpyrifos
The method and device of product.
Background technology
Chlopyrifos belongs to a kind of broad-spectrum organic insecticide for being widely used in a variety of prevention and control of plant diseases, pest control of crops.Chlopyrifos
It is the leading pesticide species of China's biocides market, output and usage amount in the world also remain high.Chlopyrifos can lead to
Cross the exposure of the media such as food, air, soil and enter human body.
Chlopyrifos can suppress cholinester activity, and the acetylcholinesterase of central nervous system is to maintaining central nervous system
The generation of nerve cell and maturation play an important roll.Increasing research confirms chlopyrifos to budding central nervous system
There is chronic toxic effect in system, defect obstacle more dynamic to children's attention, failure of memory, cognitive disorder etc. are related.America and Europe etc. sends out
Up to country highest attention is given on chlorpyrifos pollution on the intelligence of children and influence of growing.The health that environmental pollution is brought
Risk is exactly focus of concern all the time, and exposure level is to determine the pass of environmental contaminants body burden concentration in human body
Key index, and most direct index.Some organic pollutants can be metabolized after entering human body, produce a variety of metabolites,
The interior exposure of accurate quantitative analysis pollutant, a variety of metabolites of the material must be determined simultaneously.
2016《Occupational health and emergency management and rescue》One kind is described in the 4th 278-281 pages of the phase of volume 34 and uses gas phase color
The method of 3,5,6- tri- Chloro-2-Pyridyle alcohol in spectrometry measure blood plasma, it is 3 μ that this law detects lower limit to 3,5,6- tri- Chloro-2-Pyridyle alcohol
g/L.2015《Physical and chemical inspection-chemistry fascicle》One kind is described in the 9th 927-931 pages of the phase of volume 51 to connect using gas-chromatography
Mass spectrography is to the analysis methods of in the flesh of fish 3,5,6- tri- Chloro-2-Pyridyle alcohol, and the range of linearity is 1.0~1000 μ g/L, Monitoring lower-cut
0.3μg/kg.2015《Environmental Pollution and Control》One kind is described in the 9th 80-83 pages of the phase of volume 37 using double (trimethyl silicanes
Alkyl) trifluoroacetamide performs the derivatization, the method for determining chlopyrifos and its metabolite, mainly using solid phase extraction concentration,
The method that 3,5,6- tri- Chloro-2-Pyridyle alcohol in water sample are determined using gas chromatography combined with mass spectrometry method, detection are limited to 0.100 μ g/
L.2016《Chinese Professional medical science》One kind is described in the 5th 590-596 pages of the phase of volume 43 using ultra performance liquid chromatography to blood
The method that 3,5,6- tri- Chloro-2-Pyridyle alcohol are detected in slurry, the range of linearity are 1.00~50.00mg/L, and detection is limited to
0.30mg/L.2013《Chromatogram》One kind is described in the 9th 903-907 pages of the phase of volume 31 and utilizes ultra performance liquid chromatography-series connection
The method of 3,5,6- tri- Chloro-2-Pyridyle alcohol in mass spectrometric determination human urine, method detection are limited to 0.41 μ g/L.
In summary, existing document report is mainly for the chlopyrifos metabolite in blood plasma, water sample, urine death by poisoning
The detection method research of tick metabolite is less, and is concerned only with 3, a kind of 5,6- tri- metabolites of Chloro-2-Pyridyle alcohol.It is however, malicious
After dead tick enters human body, metabolism occurs in vivo and produces outside 3,5,6- tri- Chloro-2-Pyridyle alcohol, can also produce diethyl phosphate,
2 kinds of important metabolites such as diethyl thiophosphate, while rapidly and accurately quantitative determine the 3 kinds of metabolism productions of urine Chlorpyrifos
Thing is the accurate precondition for weighing exposure in chlopyrifos human body, has important work for the body burden of accurate quantitative analysis chlopyrifos
With.
And used in conventional method gas-chromatography-flame ionization detector, gas-chromatography-electron capture detection
Device, gas-chromatography-nitrogen phosphorous detector, HPLC- UV detection device etc., for 3 kinds of chlopyrifos metabolism productions of detection simultaneously
Object space face, in face of background is complicated, interfering material is more when, can not accurate qualitative, quantitative, or even there are false positive results, much without
Research and application requirement of the method to trace chlopyrifos metabolite in human sample (such as urine, blood plasma).
The content of the invention
Present invention solves the technical problem that it is the method for current human's urine Chlorpyrifos metabolite while detection
Scarcity, the present invention provide a kind of detection method of 3 kinds of main metabolites of simultaneous quantitative urine Chlorpyrifos, establish chlopyrifos
The enzymolysis of 3 kinds of metabolite urine sample detections, extraction, the method for purification and concentration, while establish and optimize 3 kinds of main generations
Thank product while the high performance liquid chromatography and second order mses (MS/MS) detection method of detection.
The technical scheme is that：
Method that is a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos, comprises the following steps：
(A) 3 kinds of metabolites of target compound selection chlopyrifos：The Chloro-2-Pyridyle alcohol of 3,5,6- tri-, diethyl phosphate,
Diethyl thiophosphate；
(B) digest：Take frost urine sample slowly to thaw at room temperature, take 10mL urine samples in 50mL centrifuge tubes, add 4mL
HCl (6M) is vibrated, and 2h is hydrolyzed under 80 DEG C of water bath conditions, is taken out after 2h, is down to room temperature；
(C) extract：10mL acetonitrile solutions are added in urine sample, after 3g NaCl, vortex mixed 1min, centrifuges, takes upper strata
5mL acetonitriles；
(D) purification and concentration：Acetonitrile after extraction is crossed into self-control purification pillar, after loading, then washed with 5mL acetonitriles
It is de-, all eluents are collected, rotary evaporated to dryness, after nitrogen drying, 1mL acetonitrile constant volumes, testing sample is in -20 DEG C of environment
It is kept in dark place, internal standard phosphoric acid di-n-butyl is added before testing；
(E) testing sample using chromatogram-from tandem mass spectrum detect, chromatogram selection Ultra Performance Liquid Chromatography instrument, chromatostrip
Part：Chromatographic column selects anti-phase C18 chromatographic columns, and column temperature is 35 DEG C, and mobile phase A is 0.2 ‰ ammoniacal liquor, and Mobile phase B is acetonitrile, sample introduction body
Product is 2 μ L, Mass Spectrometry Conditions：Ionization mode electric spray ion source, negative ion mode, more reactive ions monitor (MRM), ion gun temperature
Degree：500 DEG C, ion spray voltage：- 4500V, gas curtain gas (curtain gas, CUR) pressure is 35psi, spraying gas (ion
Source gas 1, GS1) pressure is 50psi, auxiliary heating gas (ion source gas 2, GS2) pressure is 55psi.
Further, in such scheme, the centrifugal speed in the step (C) is 8000r/min.
Further, in such scheme, in the step (D) self-control purification pillar formula for 100mg PSA,
50mg GCB, 2g anhydrous Nas2SO4Filling.
Further, in such scheme, the flow rate of mobile phase in the step (E) is 0.3mL/min, gradient elution journey
Sequence is：
| Time (min) | A (%) | B (%) |
| 0.00 | 99.0 | 1.0 |
| 2.00 | 99.0 | 1.0 |
| 3.00 | 70.0 | 30.0 |
| 10.0 | 5.0 | 95.0 |
| 13.0 | 5.0 | 95.0 |
| 13.1 | 99.0 | 1.0 |
| 15.0 | 99.0 | 1.0 |
Further, in such scheme, the selection characteristic ion in the step (E) is to being respectively：3,5,6- tri- is chloro-
2- pyridine alcohol：(196.0/35.0 m/z) and 198.0/35.0 (m/z)；Diethyl phosphate：(153.0/79.0 m/z) and 153.0/
125.0(m/z)；Diethyl thiophosphate：(169.0/95.0 m/z) and 169.0/141.0 (m/z).
Select ion parameters for：
Device used in described method that is a kind of while detecting urine Chlorpyrifos kind metabolite mainly includes：Main body
And detecting instrument, the detecting instrument are ultra performance liquid chromatography GC-MS；Supply unit, defrosting are included in the main body
Device, centrifugal device one, water bath device, vortex mixed device, centrifugal device two, purifier, rotary evaporating device, PLC controls
Device processed, the supply unit are located at the left side of the main body, the thawing apparatus, positioned at the back upper place of the supply unit, institute
State centrifugal device one, centrifugal device two, purifier, rotary evaporating device and be located at body interior, the water from left to right successively
Bath apparatus is located at the front of centrifugal device one, and the vortex mixed device is located at the front of centrifugal device two, the PLC
Positioned at the upper right side of main body, the rear of the thawing apparatus is provided with Hl storage devices, main body back side top from a left side to
The right side is sequentially provided with acetonitrile storage device, nitrogen storage device, internal standard phosphoric acid di-n-butyl storage device, and the thawing apparatus passes through
Conduit one is connected to the centrifugal device one, and flow control valve one is provided with the conduit one, and the Hl storage devices pass through
Conduit two is connected to the centrifugal device one, and flow control valve two is provided with the conduit two, and the centrifugal device one passes through
Conduit three is connected to the water bath device, and flow control valve three is provided with the conduit three, and the water bath device passes through conduit
Four are connected to the vortex mixed device, and flow control valve four is provided with the conduit four, and the vortex mixed device passes through
Conduit five is connected to the acetonitrile storage device, and flow control valve five, the vortex mixed device are provided with the conduit five
Conduit six is connected to the centrifugal device two, and flow control valve six is provided with the conduit six, and the centrifugal device two passes through
Conduit seven is connected to the purifier, and the conduit seven is provided with flow control valve seven, and the purifier passes through conduit eight
The acetonitrile storage device is connected to, flow control valve eight is provided with the conduit eight, the rotary evaporating device is provided with
Revolving bottle, receiving flask, connector one and connector two, the purifier are connected to the rotary evaporating device by conduit nine
Connector one, be provided with flow control valve nine on the conduit nine, the connector two on the rotary evaporating device is by leading
Pipe ten is connected to the nitrogen storage device, and flow control valve ten is provided with the conduit ten, and the receiving flask passes through conduit
11 are connected to the internal standard phosphoric acid di-n-butyl storage device, and flow control valve 11, institute are provided with the conduit 11
State flow control valve one, flow control valve two, flow control valve three, flow control valve four, flow control valve five, flow control valve
6th, flow control valve seven, flow control valve eight, flow control valve nine, flow control valve ten, flow control valve 11 are by each
From electromagnetism relay valve and wire be connected to the PLC, the mark phosphoric acid di-n-butyl storage device passes through catheter
The chromatogram-from tandem mass spectrum detector is connected to, the supply unit is the thawing apparatus, centrifugal device one, water-bath dress
Put, vortex mixed device, centrifugal device two, purifier, rotary evaporating device, PLC provide power supply.The present apparatus is very
Suitable for the method for the present invention, the two is very good with the use of effect.
Further, in such scheme, heater, the bottom of centrifugal device one are provided with the outside of the thawing apparatus
Provided with electric rotating machine one, the water bath device bottom is provided with water-bath heater, and the vortex mixed bottom of device is provided with electronic mixed
Clutch, the bottom of centrifugal device two are provided with electric rotating machine two, the heater, electric rotating machine one, water-bath heater, electronic
Blender, electric rotating machine two are connected to the supply unit, and control panel is provided with the supply unit.The PLC controls
Device processed is by by innernal CPU, the unit institute modularity such as instruction and data internal memory, input-output unit, power supply module, digital simulation
It is combined into.
The present invention principle be：By being digested, extracting, purifying and being concentrated to urine sample, to urine Chlorpyrifos 3
Kind main metabolites carry out Extraction and enrichment, and self-control purification pillar significantly reduces the interference such as pigment in urine, sugar, salinity；It is logical
Cross and quantitative, qualitative analysis, the series methods are carried out to target compound using two level Four mass analyzers using tandem mass spectrum
Anti-background interference ability is strong, qualitative reliable, quantitative accurate.
As a result：(3 times of the instrument test limit of HPLC-MS/MS detection methods after optimization to 3 kinds of metabolites of chlopyrifos
Signal to noise ratio) to be 0.001mg/L, quantitative test limit is 0.005mg/L (10 times of signal to noise ratio), 5 standard curve phase relations
Number (R2) between 0.9916-0.9991, the recovery of standard addition of urine Chlorpyrifos is 80.3%~90.0%, and it is fixed to disclosure satisfy that
Measure the requirement of analysis.Its rate of recovery result is as follows：The Chloro-2-Pyridyle alcohol of 3,5,6- tri-, diethyl phosphate, diethyl thiophosphate；
The beneficial effects of the invention are as follows：The invention provides a kind of while detect 3 kinds of metabolites of urine Chlorpyrifos
Liquid phase chromatogram-mass spectrometry combination method and device, this method be by urine sample is digested, extract, purify with concentration, it is right
3 kinds of main metabolites of urine Chlorpyrifos carry out Extraction and enrichments, self-control purification pillar significantly reduce pigment in urine, sugar,
Salinity etc. disturbs；Using the multiple-reaction monitoring technology (MRM) of high performance liquid chromatography/tandem mass spectrum (HPLC-MS/MS) come solve with
The selectivity of trace target components, sensitivity in detection of complex background matrix toward conventional detector such as UV-Vis, ECD, NPD
Present on inferior position, enormously simplify sample pretreatment step using the background decrease of noise functions of second order mses, improve sample and determine
Property accuracy, due to MRM technologies can have concurrently sensitivity and it is selective the advantages of, qualitative results accurately and reliably, while accurate are surveyed
Determine 3 kinds of main metabolites of urine Chlorpyrifos.
There is the advantages of succinct quick using the device of the present invention, solve in existing laboratory that equipment is not complete, or
The problem of operation sequence complexity, can disposable rapid results, and realize intelligentized control method, controllability is strong, accurately
Degree is high, reduces error, greatlys save the time.
The more conventional detection method of method after being optimized using the present invention has preferably selectivity and higher sensitivity, such a
There is significant advantage on second order mses detection technique 3 kinds of metabolites in chlopyrifos human urine are qualitative, can be by simple
Quick batch pre-treating method, the demand of Quantitative detection urine 3 kinds of main metabolites concentration of Chlorpyrifos, is carried significantly
The high validity and reliability of detection, saves detection time and testing cost.
Brief description of the drawings
Fig. 1 is the structural representation of equipment therefor of the present invention；
Fig. 2 is 3 kinds of metabolites of chlopyrifos and internal standard reference colour spectrogram (the Chloro-2-Pyridyle alcohol of 3,5,6- tri-：TCPyr, phosphoric acid
Diethylester：DEP, diethyl thiophosphate：DETP, phosphoric acid di-n-butyl：DBP)；
Fig. 3 is blank urine sample chromatogram；
Fig. 4 is mark-on urine sample chromatogram (the Chloro-2-Pyridyle alcohol of 3,5,6- tri-：TCPyr, diethyl phosphate：DEP, D2EHDTPA
Diethylester：DETP, phosphoric acid di-n-butyl：DBP)；
Fig. 5 is actual urine sample typical color spectrogram (the Chloro-2-Pyridyle alcohol of 3,5,6- tri-：TCPyr, diethyl phosphate：It is DEP, thio
Diethyl phosphate：DETP, phosphoric acid di-n-butyl：DBP).
Wherein, 1- main bodys, 2- supply units, 3- thawing apparatus, 4- centrifugal devices one, 5- water bath devices, 6- vortex mixeds
Device, 7- centrifugal devices two, 8- purifiers, 9- rotary evaporating devices, 10-PLC controllers, 11-HCl storage devices, 12- second
Nitrile storage device, 13- nitrogen storage device, 14- internal standard phosphoric acid di-n-butyls storage device, 15- catheters, 16- conduits one,
17- conduits two, 18- conduits three, 19- conduits four, 20- conduits five, 21- conduits six, 22- conduits seven, 23- conduits eight, 24- conduits
9th, 25- conduits ten, 26- conduits 11,27- flow control valves one, 28- flow control valves two, 29- flow control valves three, 30-
Flow control valve four, 31- flow control valves five, 32- flow control valves six, 33- flow control valves seven, 34- flow control valves eight,
35- flow control valves nine, 36- flow control valves ten, 37- flow control valves 11,38- wires, 2a- control panels, 3a- heating
Device, 4a- electric rotating machines one, 5a- water-bath heaters, 6a- motorized agitators, 7a- electric rotating machines two, 9a- revolving bottles, 9b- are received
Collect bottle, 9c- connectors one, 9d- connectors two.
Embodiment
By the following examples the present invention is further illustrated with reference to accompanying drawing.Protection scope of the present invention is not restricted to
The present embodiment.
Embodiment 1：
(1) standard curve is prepared：
Accurately weigh chlopyrifos metabolite standard specimen and internal standard (DBP) (Dr.Ehrenstorfer GmbH, Germany)
0.01 ± 0.0001g, with acetonitrile dissolving and constant volume, obtains TCPyr, DEP, DETP that concentration is 1000mg/L into 10mL volumetric flasks
And internal standard DBP Standard Stock solutions.And with dilution in acetonitrile, preparation obtain concentration for 0.005,0.01,0.02,0.05,0.10,
0.20mg/L working solution.
(2) instrumental conditions：
Instrument chromatographic condition：The ultra performance liquid chromatography is the 1290Infinity of Agilent companies；Chromatographic column selects
The anti-phase C18 chromatographic columns of Agilent (3.5 μm of ZORBAX Eclipse Plus C18,2.1 × 150mm, Agilent), column temperature is
35 DEG C, mobile phase A is 0.2 ‰ ammoniacal liquor, and Mobile phase B is acetonitrile, and flow velocity 0.3mL/min, sampling volume is 2 μ L；Mobile phase ladder
Spend and be：
| Time (min) | A (%) | B (%) |
| 0.00 | 99.0 | 1.0 |
| 2.00 | 99.0 | 1.0 |
| 3.00 | 70.0 | 30.0 |
| 10.0 | 5.0 | 95.0 |
| 13.0 | 5.0 | 95.0 |
| 13.1 | 99.0 | 1.0 |
| 15.0 | 99.0 | 1.0 |
Mass Spectrometry Conditions：Ionization mode electric spray ion source, negative ion mode, more reactive ions monitor (MRM), ion gun temperature
Degree：500 DEG C, ion spray voltage：- 4500V, gas curtain gas (curtain gas, CUR) pressure is 35psi, spraying gas (ion
Source gas 1, GS1) pressure is 50psi, auxiliary heating gas (ion source gas 2, GS2) pressure is 55psi.Selection
Ion parameters are：
(3) Specification Curve of Increasing：
Sample introduction is analyzed under above-mentioned chromatographic condition, 3,5,6- tri- Chloro-2-Pyridyle alcohol, diethyl phosphate and D2EHDTPA diethyl
The retention time of ester, linear equation coefficient correlation.Y in linear equation：Represent the peak area and internal standard phosphoric acid of chlopyrifos metabolin
The peak area ratio of di-n-butyl.
And thereby determine that, the instrument detection limit (LOD) of 3 kinds of chlopyrifos metabolites is 0.001mg/L, and instrument quantitative limits
(LOQ) it is 0.005mg/L.
Embodiment 2：
Method that is a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos, comprises the following steps：
(A) 3 kinds of metabolites of target compound selection chlopyrifos：The Chloro-2-Pyridyle alcohol of 3,5,6- tri-, diethyl phosphate,
Diethyl thiophosphate；
(B) 6 parts of blank urine samples are weighed, every part of urine samples 10.0mL (± 0.01mL), two groups is divided into, adds respectively
0.01mg/kg, 0.050mg/kg, 0.20mg/L chlopyrifos metabolite standard liquid, by above-mentioned sample-pretreating method：Add
Enter 4mL HCl (6M) vibrations, 2h is hydrolyzed under 80 DEG C of water bath conditions.Taken out after 2h, be down to room temperature.
(C) extract：10mL acetonitrile solutions are added in urine sample, after 3g NaCl, vortex mixed 1min, are centrifuged with 8000r/min
Separation, takes upper strata 5mL acetonitriles；
(D) purification and concentration：Acetonitrile after extraction is crossed into self-control purification pillar, the formula of self-control purification pillar is
100mg PSA, 50mg GCB, 2g anhydrous Nas2SO4Filling, after loading, then eluted with 5mL acetonitriles, collect all eluents,
Rotary evaporated to dryness, after nitrogen drying, 1mL acetonitrile constant volumes, testing sample is kept in dark place in -20 DEG C of environment, adds before test
Enter internal standard phosphoric acid di-n-butyl；
(E) instrumental conditions：
Instrument chromatographic condition：The ultra performance liquid chromatography is the 1290Infinity of Agilent companies；Chromatographic column selects
The anti-phase C18 chromatographic columns of Agilent (3.5 μm of ZORBAX Eclipse Plus C18,2.1 × 150mm, Agilent), column temperature is
35 DEG C, mobile phase A is 0.2 ‰ ammoniacal liquor, and Mobile phase B is acetonitrile, and flow velocity 0.3mL/min, sampling volume is 2 μ L；Mobile phase ladder
Spend and be：
| Time (min) | A (%) | B (%) |
| 0.00 | 99.0 | 1.0 |
| 2.00 | 99.0 | 1.0 |
| 3.00 | 70.0 | 30.0 |
| 10.0 | 5.0 | 95.0 |
| 13.0 | 5.0 | 95.0 |
| 13.1 | 99.0 | 1.0 |
| 15.0 | 99.0 | 1.0 |
Mass Spectrometry Conditions：Ionization mode electric spray ion source, negative ion mode, more reactive ions monitor (MRM), ion gun temperature
Degree：500 DEG C, ion spray voltage：- 4500V, gas curtain gas (curtain gas, CUR) pressure is 35psi, spraying gas (ion
Source gas 1, GS1) pressure is 50psi, auxiliary heating gas (ion source gas 2, GS2) pressure is 55psi.Selection
Ion parameters are：
As a result show, when urine sample Chlorpyrifos metabolite addition concentration is 0.01~0.2mg/kg, the rate of recovery is
80.3%~90..0%, relative standard deviation 1.20%~6.47%.
Embodiment 3：
(A) target compound is the Chloro-2-Pyridyle alcohol of 3,5,6- tri-, diethyl phosphate and diethyl thiophosphate；
(B) sample collection：Sampling time is September in 2016 2, and sampling position is Nanjing Gaochun area Ya Xi towns, tested
Urine sample comes from children；
(C) digest：Take frost urine sample slowly to thaw at room temperature, take 10mL urine samples in 50mL centrifuge tubes, add 4mL
HCl (6M) is vibrated, and 2h is hydrolyzed under 80 DEG C of water bath conditions, is taken out after 2h, is down to room temperature；
(D) extract：10mL acetonitrile solutions are added in urine sample, after 3g NaCl, vortex mixed 1min, after centrifugation, are taken
Layer 5mL acetonitriles；
(E) purification and concentration：Acetonitrile after extraction is crossed into self-control purification pillar, after loading, then washed with 5mL acetonitriles
It is de-, all eluents are collected, rotary evaporation is done near, after nitrogen drying, 1mL acetonitrile constant volumes, environment of the testing sample at -20 DEG C
In be kept in dark place, add internal standard phosphoric acid di-n-butyl before testing, sample is measured using high performance liquid chromatography tandem mass spectrum method,
Condition determination is as follows：
Instrument chromatographic condition：The ultra performance liquid chromatography is the 1290Infinity of Agilent companies；Chromatographic column selects
The anti-phase C18 chromatographic columns of Agilent (3.5 μm of ZORBAX Eclipse Plus C18,2.1 × 150mm, Agilent), column temperature is
35 DEG C, mobile phase A is 0.2 ‰ ammoniacal liquor, and Mobile phase B is acetonitrile, and flow velocity 0.3mL/min, sampling volume is 2 μ L；Mobile phase ladder
Spend and be：
| Time (min) | A (%) | B (%) |
| 0.00 | 99.0 | 1.0 |
| 2.00 | 99.0 | 1.0 |
| 3.00 | 70.0 | 30.0 |
| 10.0 | 5.0 | 95.0 |
| 13.0 | 5.0 | 95.0 |
| 13.1 | 99.0 | 1.0 |
| 15.0 | 99.0 | 1.0 |
Mass Spectrometry Conditions：Ionization mode electric spray ion source, negative ion mode, more reactive ions monitor (MRM), ion gun temperature
Degree：500 DEG C, ion spray voltage：- 4500V, gas curtain gas (curtain gas, CUR) pressure is 35psi, spraying gas (ion
Source gas 1, GS1) pressure is 50psi, auxiliary heating gas (ion source gas 2, GS2) pressure is 55psi.Selection
Ion parameters are：
Testing result shows, 3,5,6- tri- Chloro-2-Pyridyle determining alcohols are 15.41 μ g/g in children's urine, p diethylaminobenzoic acid
Ester concentration is 7.62 μ g/g, and diethyl thiophosphate concentration is 4.36 μ g/g.
Device used in above-described embodiment 1-3 mainly includes：Main body 1 and detecting instrument 39, detecting instrument 39 are ultra high efficiency
Liquid chromatography mass combined instrument, the QTRAP 4500 of the 1290Infinity+AB Sciex companies selected from Agilent companies are main
In body 1 comprising supply unit 2, thawing apparatus 3, centrifugal device 1, water bath device 5, vortex mixed device 6, centrifugal device 27,
Purifier 8, rotary evaporating device 9, PLC 10, supply unit 2 are located at the left side of main body 1, thawing apparatus 3, positioned at electricity
The back upper place of source device 2, centrifugal device 1, centrifugal device 27, purifier 8, rotary evaporating device 9 position from left to right successively
Inside main body 1, water bath device 5 is located at the front of centrifugal device 1, before vortex mixed device 6 is located at centrifugal device 27
Side, PLC 10 are located at the upper right side of main body 1, the rear of thawing apparatus 3 are provided with HCl storage devices 11, in main body 1
Back side top be sequentially provided with from left to right acetonitrile storage device 12, nitrogen storage device 13, internal standard phosphoric acid di-n-butyl storage
Device 14, thawing apparatus 3 are connected to centrifugal device 1 by conduit 1, and flow control valve 1 is provided with conduit 1,
HCl storage devices 11 are connected to centrifugal device 1 by conduit 2 17, and flow control valve 2 28 is provided with conduit 2 17, from
Center device 1 is connected to water bath device 5 by conduit 3 18, and flow control valve 3 29, water bath device 5 are provided with conduit 3 18
Vortex mixed device 6 is connected to by conduit 4 19, flow control valve 4 30, vortex mixed device 6 are provided with conduit 4 19
Acetonitrile storage device 12 is connected to by conduit 5 20, flow control valve 5 31, vortex mixed device 6 are provided with conduit 5 20
Conduit 6 21 is connected to centrifugal device 27, and flow control valve 6 32 is provided with conduit 6 21, and centrifugal device 27 passes through conduit
7 22 are connected to purifier 8, and conduit 7 22 is provided with flow control valve 7 33, and purifier 8 is connected to by conduit 8 23
Acetonitrile storage device 12, is provided with flow control valve 8 34 on conduit 8 23, and rotary evaporating device 9 is provided with revolving bottle 9a, received
Collection bottle 9b, the 9c of connector one and the 9d of connector two, purifier 8 are connected to the connection of rotary evaporating device 9 by conduit 9 24
A mouthful 9c, is provided with flow control valve 9 35 on conduit 9 24, and the 9d of connector two on rotary evaporating device 9 passes through conduit 10
Nitrogen storage device 13 is connected to, flow control valve 10 is provided with conduit 10, receiving flask 9b is connected by conduit 11
Internal standard phosphoric acid di-n-butyl storage device 14 is connected to, flow control valve 11, flow control valve are provided with conduit 11
One 27, flow control valve 2 28, flow control valve 3 29, flow control valve 4 30, flow control valve 5 31, flow control valve six
32nd, flow control valve 7 33, flow control valve 8 34, flow control valve 9 35, flow control valve 10, flow control valve 11
37 are connected to PLC 10 by respective electromagnetism relay valve and wire 38, and mark phosphoric acid di-n-butyl storage device 14 is logical
Cross catheter 15 and be connected to detecting instrument 39, supply unit 2 be thawing apparatus 3, centrifugal device 1, water bath device 5, be vortexed it is mixed
Attach together and 6, centrifugal device 27, purifier 8, rotary evaporating device 9, the offer power supply of PLC 10 are provided.The outside of thawing apparatus 3
Provided with heater 3a, the bottom of centrifugal device 1 is provided with the 4a of electric rotating machine one, and the bottom of water bath device 5 is provided with water-bath heater 5a,
The bottom of vortex mixed device 6 is provided with motorized agitator 6a, the bottom of centrifugal device 27 be provided with electric rotating machine two 7a, heater 3a,
The 4a of electric rotating machine one, water-bath heater 5a, motorized agitator 6a, the 7a of electric rotating machine two are connected to supply unit 2, are filled in power supply
Put 2 and be provided with control panel 2a.
The Chloro-2-Pyridyle alcohol of 3,5,6- tri-, diethyl phosphate, the rate of recovery result of diethyl thiophosphate are as follows：
As a result show：The instrument test limit of HPLC-MS/MS detection methods after optimization to 3 kinds of metabolites of chlopyrifos
(3 times of signal to noise ratio) to be 0.001mg/L, quantitative test limit is 0.005mg/L (10 times of signal to noise ratio), 5 standard curve phases
Relation number (R2) between 0.9916-0.9991, the recovery of standard addition of urine Chlorpyrifos is 80.3%~90.0%, Neng Gouman
The requirement of sufficient quantitative analysis.
Finally it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；To the greatest extent
The present invention is described in detail with reference to the foregoing embodiments for pipe, it will be understood by those within the art that：It is still
Technical scheme described in previous embodiment can be modified, or equivalent substitution is carried out to which part technical characteristic；
And these modifications or replacement, the essence of appropriate technical solution is departed from the spirit and model of technical scheme of the embodiment of the present invention
Enclose.
Claims (8)
1. method that is a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos, it is characterised in that comprise the following steps：
(A) 3 kinds of interior metabolism products of target compound selection chlopyrifos：The Chloro-2-Pyridyle alcohol of 3,5,6- tri-, diethyl phosphate,
Diethyl thiophosphate；
(B) digest：Take frost urine sample slowly to thaw at room temperature, take 10mL urine samples in 50mL centrifuge tubes, add 4mL HCl
(6M) is vibrated, and 2h is hydrolyzed under 80 DEG C of water bath conditions, is taken out after 2h, is down to room temperature；
(C) extract：10mL acetonitrile solutions are added in urine sample, after 3g NaCl, vortex mixed 1min, centrifuges, takes upper strata 5mL
Acetonitrile；
(D) purification and concentration：Acetonitrile after extraction is crossed and makes purification pillar by oneself, after loading, then with the elution of 5mL acetonitriles, is received
Collect all eluents, rotary evaporated to dryness, after nitrogen drying, 1mL acetonitrile constant volumes, testing sample lucifuge in -20 DEG C of environment
Preserve, internal standard phosphoric acid di-n-butyl is added before testing；
(E) testing sample using chromatogram-from tandem mass spectrum detect, chromatogram selection Ultra Performance Liquid Chromatography instrument, chromatographic condition：Color
Compose post and select anti-phase C18 chromatographic columns, column temperature is 35 DEG C, and mobile phase A is 0.2 ‰ ammoniacal liquor, and Mobile phase B is acetonitrile, sampling volume 2
μ L, Mass Spectrometry Conditions：Ionization mode electric spray ion source, negative ion mode, more reactive ions monitor (MRM), ion source temperature：
500 DEG C, ion spray voltage：- 4500V, gas curtain gas (curtain gas, CUR) pressure is 35psi, spraying gas (ion
Source gas 1, GS1) pressure is 50psi, auxiliary heating gas (ion source gas 2, GS2) pressure is 55psi.
2. method that is according to claim 1 a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos, its feature exist
In the centrifugal speed in the step (C) is 8000r/min.
3. according to a kind of described in claim 1 while the method that detects 3 kinds of metabolites of urine Chlorpyrifos, its feature exists
In the formula of the self-control purification pillar in the step (D) is 100mg PSA, 50mg GCB, 2g anhydrous Nas2SO4Filling.
4. according to a kind of described in claim 1 while the method that detects 3 kinds of metabolites of urine Chlorpyrifos, its feature exists
In the flow rate of mobile phase in the step (E) is 0.3mL/min or so, and gradient elution program is 0~2min, 99%A:1%B,
2~3min, 70%A:30%B, 3~10min, 5%A:95%B, 10~13min, 5%A:95%B, 13.1~15min, 99%
A:1%B.
5. according to a kind of described in claim 1 while the method that detects 3 kinds of metabolites of urine Chlorpyrifos, its feature exists
In the gradient elution program in the step (E) is 0~2min, 99%A:1%B, 2~3min, 70%A:30%B, 3~
10min, 5%A:95%B, 10~13min, 5%A:95%B, 13.1~15min, 99%A:1%B.
6. according to a kind of described in claim 4 while the method that detects 3 kinds of metabolites of urine Chlorpyrifos, its feature exists
In the selection characteristic ion in the step (E) is to being respectively：The Chloro-2-Pyridyle alcohol of 3,5,6- tri-：196.0/35.0 (m/z) and
198.0/35.0(m/z)；Diethyl phosphate：(153.0/79.0 m/z) and 153.0/125.0 (m/z)；Diethyl thiophosphate：
(169.0/95.0 m/z) and 169.0/141.0 (m/z).
7. a kind of according to claim 1-6 any one while the method for detecting 3 kinds of metabolites of urine Chlorpyrifos
Device used, it is characterised in that mainly include：Main body (1) and detecting instrument (39), the detecting instrument (39) are ultra high efficiency
Liquid chromatography mass combined instrument；Supply unit (2), thawing apparatus (3), centrifugal device one (4), water are included in the main body (1)
Bath apparatus (5), vortex mixed device (6), centrifugal device two (7), purifier (8), rotary evaporating device (9), PLC
(10), the supply unit (2) is located at the left side of the main body (1), the thawing apparatus (3), positioned at the supply unit (2)
Back upper place, the centrifugal device one (4), centrifugal device two (7), purifier (8), rotary evaporating device (9) are successively from a left side
Internal positioned at main body (1) to the right side, the water bath device (5) is located at the front of centrifugal device one (4), the vortex mixed device
(6) it is located at the front of centrifugal device two (7), the PLC (10) is located at the upper right side of main body (1), in the dress that thaws
The rear for putting (3) is provided with HCl storage devices (11), and acetonitrile storage is sequentially provided with from left to right in the back side top of main body (1)
Device (12), nitrogen storage device (13), internal standard phosphoric acid di-n-butyl storage device (14), the thawing apparatus (3) is by leading
Pipe one (16) is connected to the centrifugal device one (4), and flow control valve one (27) is provided with the conduit one (16), described
HCl storage devices (11) are connected to the centrifugal device one (4) by conduit two (17), and described (4) bottom of centrifugal device one is set
There is electric rotating machine one (4a), flow control valve two (28) is provided with the conduit two (17), the centrifugal device one (4) passes through
Conduit three (18) is connected to the water bath device (5), and flow control valve three (29), the water are provided with the conduit three (18)
Bath apparatus (5) is connected to the vortex mixed device (6) by conduit four (19), and flow control is provided with the conduit four (19)
Valve four (30) processed, the vortex mixed device (6) is connected to the acetonitrile storage device (12) by conduit five (20), described
Conduit five (20) is provided with flow control valve five (31), and vortex mixed device (6) conduit six (21) is connected to the centrifugation
Device two (7), is provided with flow control valve six (32) on the conduit six (21), and the centrifugal device two (7) passes through conduit seven
(22) purifier (8) is connected to, the conduit seven (22) is provided with flow control valve seven (33), the purifier
(8) the acetonitrile storage device (12) is connected to by conduit eight (23), flow control valve is provided with the conduit eight (23)
Eight (34), the rotary evaporating device (9) are provided with revolving bottle (9a), receiving flask (9b), connector one (9c) and connector two
(9d), the purifier (8) are connected to the connector one (9c) of the rotary evaporating device (9) by conduit nine (24),
The conduit nine (24) is provided with flow control valve nine (35), and the connector two (9d) on the rotary evaporating device (9) passes through
Conduit ten (25) is connected to the nitrogen storage device (13), and flow control valve ten (36) is provided with the conduit ten (25),
The receiving flask (9b) is connected to the internal standard phosphoric acid di-n-butyl storage device (14) by conduit 11 (26), is led described
Guan Shiyi (26) is provided with flow control valve 11 (37), the flow control valve one (27), flow control valve two (28), flow
Control valve three (29), flow control valve four (30), flow control valve five (31), flow control valve six (32), flow control valve seven
(33), flow control valve eight (34), flow control valve nine (35), flow control valve ten (36), flow control valve 11 (37) are equal
The PLC (10), the mark phosphoric acid di-n-butyl storage are connected to by respective electromagnetism relay valve and wire (38)
Device (14) is connected to the detecting instrument (39) by catheter (15), and the supply unit (2) is the thawing apparatus
(3), centrifugal device one (4), water bath device (5), vortex mixed device (6), centrifugal device two (7), purifier (8), rotation
Vaporising device (9), PLC (10) provide power supply.
8. the device used in method that is according to claim 7 a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos,
Characterized in that, being provided with heater (3a) on the outside of the thawing apparatus (3), described (4) bottom of centrifugal device one is provided with rotation
Motor one (4a), water bath device (5) bottom are provided with water-bath heater (5a), and vortex mixed device (6) bottom is provided with
Motorized agitator (6a), described (7) bottom of centrifugal device two are provided with electric rotating machine two (7a), the heater (3a), rotation
Motor one (4a), water-bath heater (5a), motorized agitator (6a), electric rotating machine two (7a) are connected to the supply unit
(2) control panel (2a), is provided with the supply unit (2).
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| CN108717094A (en) * | 2018-05-08 | 2018-10-30 | 杭州谱可检测技术有限公司 | The device and method of l-cn in detection baby milk powder based on LC-MS |
| CN109507335A (en) * | 2018-12-29 | 2019-03-22 | 上海交通大学医学院附属新华医院 | Utilize the method for a variety of different type environmental contaminants in LC-MS-MS high throughput detection urine |
| CN114813991A (en) * | 2022-03-09 | 2022-07-29 | 上海交通大学医学院 | Method for detecting neonicotinoid insecticide in human urine |
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