CN109507335A - Utilize the method for a variety of different type environmental contaminants in LC-MS-MS high throughput detection urine - Google Patents
Utilize the method for a variety of different type environmental contaminants in LC-MS-MS high throughput detection urine Download PDFInfo
- Publication number
- CN109507335A CN109507335A CN201811633115.9A CN201811633115A CN109507335A CN 109507335 A CN109507335 A CN 109507335A CN 201811633115 A CN201811633115 A CN 201811633115A CN 109507335 A CN109507335 A CN 109507335A
- Authority
- CN
- China
- Prior art keywords
- mobile phase
- sample
- detection
- volume
- urine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid 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
- G01N2030/062—Preparation extracting sample from raw material
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Pathology (AREA)
- Physics & Mathematics (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
The present invention discloses a kind of method using a variety of different type environmental contaminants in LC-MS/MS high throughput detection urine, the following steps are included: (1) Sample pretreatment: being sufficient vortex after acetonitrile treatment of the urine containing formic acid after digesting, centrifuging and taking supernatant, traditional vacuum concentration is carried out to supernatant, the sample introduction sample of machine testing in acquisition；(2) using sample introduction sample described in LC-MS/MS sample detection: liquid-phase condition are as follows: chromatographic column uses reverse phase C18 chromatographic column；Mobile phase A is the ammonium acetate aqueous solution of 4~6mmol/L；Mobile phase B is methanol；Using gradient elution；Detect Mass Spectrometry Conditions are as follows: using the dynamic polyion detection pattern under electric spray ion source.Method of the invention has the characteristics that quick detection, high efficiency, high throughput, versatile, high sensitivity, stability and favorable reproducibility, the mass detection of biological sample suitable for epidemiological study.
Description
Technical field
The invention belongs to environmental analysis fields, and in particular to it is a kind of using LC-MS/MS high throughput detection urine in it is a variety of not
Exploitation, verifying and the application of the method for same type environmental contaminants.
Background technique
While China's economic fast development, influence of the brought environmental pollution especially to health is concerned.Closely
In the past few years find detect in general population's urine in the detection work of environmental contaminants in my be engaged in human sample
To variety classes environmental contaminants, but it is based respectively on different detection methods.Since people unavoidably subject diversification
The cumulative exposure of pollutant, rather than the exposure of Single Pollution object, how the exposure level of thoroughly evaluating human body multiple pollutant, be
Research environment pollutes the premise and basis to health effect.In the biological sample rare modern times, it is more to develop a kind of disposable detection
The method for high-flux analysis of seed type difference pollutant is the key that then carry out crowd's pollutant thoroughly evaluating.
China is that an agricultural and industrial power, pesticide and chemical products are widely used by long-term, is brought a variety of
Environmental pollution.Wherein, four class pesticides (organic phosphorus and classes of herbicides, carbamates, pyrethroid and anabasine),
Three classes anti-corrosion/bacteriostatic agent (parabens, phthalate, triclosan and triclocarban class etc.), phenols and
9 pollutants such as other chemical products (- 3 class of Benzophenone etc.), are widely present in our daily lifes, and have low dosage
The features such as duration, specificity, passage property and slowness is current domestic and international important pollutant of greatest concern, specific to represent
Compound is as shown in table 1.Persistence organic pollutant (POPs) is often under the lasting exposure of low concentration to people in these environment
Body generates certain harm, to influence human health development.Therefore, we are badly in need of existing to multiple pollutant in above-mentioned 9 major class
Exposure in crowd carries out comprehensive and accurate assessment.
The current published detection method for a variety of environmental contaminants, it is fewer to cover pollutant type, general
Property is low, and method sensitivity is lower.For example, being adopted using the method for single phenols environmental contaminants in GC-MS/MS detection urine
The method for detecting benzoates, phenols two major classes environmental contaminants in urine with LC-MS/MS, and detected using GC-MS/MS
Organophosphorus pesticide, phthalate, method of bisphenol-A (BPA) three classes environmental contaminants etc. in urine.Wherein, GC-MS
Method Sample pretreatment needs the complex steps such as acidification, liquid-liquid extraction, Solid Phase Extraction and derivatization, time and effort consuming, and GC
It is difficult to apply to the compound of heated structure destructible, using there are certain limitations.The pollutant type that LC-MS method is related to
But not enough extensively, need repeated detection so as to cause with sample amount is bigger, time-consuming ratio when detecting a variety of different types of pollutants
It is longer.Therefore, the prior art is also faced with challenge to pollutant exposure thoroughly evaluating, and urgent need develops one kind disposably with a small amount of sample
Detect the sensitive stable detection method of a variety of different type pollutants.
For liquid phase analysis, disposably detects difficulty present in a variety of different type compounds and be: in liquid phase
It is difficult to for different types of each compound to be kept completely separate, single compound sensitivity of mass spectrometry reduces etc..It develops a kind of disposable
With the sensitive stable detection method of a variety of different type pollutants of a small amount of pattern detection, there is also certain difficulties.Due to influenceization
There are also the effect of sample mesostroma other than the performance of detecting instrument itself, developing one kind can have the reason of closing object sensitivity
Effect removal sample matrix can sufficiently be enriched with the Sample pretreatment method of target compound again and LC-MS/MS is combined to become analysis
The key that method is successfully established.
Summary of the invention
Technical problem to be solved by the present invention lies in provide a variety of inhomogeneities in a kind of disposable high-throughput detection urine
The method of type environmental contaminants has quick detection, high efficiency, high throughput, versatile, high sensitivity, stability and reproduction
The good feature of property.
In order to solve the above technical problems, the technical solution adopted by the present invention are as follows:
A method of a variety of different type environmental contaminants in urine being detected using LC-MS/MS high throughput, including following
Step:
(1) it Sample pretreatment: takes the urine specimen of certain volume that internal standard compound is added, the vinegar of half urine sample volume is added
Sour ammonium/acetate buffer salt tune ph to 5.0 or so is added β glucuronic acid esterase and is incubated for a period of time；Add after incubation
Entering the acetonitrile of 0.1~2% formic acid containing volume fraction, centrifuging and taking supernatant after sufficient vortex carries out traditional vacuum concentration to supernatant,
The sample introduction sample of machine testing in acquisition；
(2) using sample introduction sample described in LC-MS/MS sample detection:
Detect liquid-phase condition are as follows: chromatographic column uses reverse phase C18 chromatographic column；Mobile phase includes mobile phase A and Mobile phase B；Stream
The ammonium acetate aqueous solution that dynamic phase A is 4~6mmol/L；Mobile phase B is methanol；Using gradient elution, gradient elution program is by following
Program carries out: mobile phase A+Mobile phase B=100%；Mobile phase original ratio is 95% mobile phase A and 5% Mobile phase B；
0~4min, Mobile phase B percent by volume are incremented to 10% by 5%；4~12min, Mobile phase B percent by volume are incremented by by 10%
To 60%；12~14min, Mobile phase B percentage by volume are incremented to 85% by 60%；14~16min, Mobile phase B volume basis
Number is incremented to 90% by 85%；16~18min, it is 90% that Mobile phase B, which keeps percentage by volume,；
Detect Mass Spectrometry Conditions are as follows: using the dynamic polyion detection pattern (DMRM) under electric spray ion source (ESI).
It in a specific embodiment, is using ammonium acetate aqueous solution tune pH in step (1).Preferably, the acetic acid
The concentration of aqueous ammonium is 1mol/L.
In a specific embodiment, in step (1), incubation conditions are 37 DEG C of 2~4h of incubation.Preferably, it is incubated for 3h.
In a specific embodiment, in step (1), the addition volume of the acetonitrile of 0.1~2% formic acid is urine specimen
3~8 times of volume.Preferably 4~7 times.More preferably 5~6 times.
In a specific embodiment, in step (1), the volume fraction of formic acid is 0.5~1.5% in the acetonitrile.
Preferably 0.8~1.2%.More preferably 0.9~1.1%.
In a specific embodiment, it is 4mmol/L, 4.5mmol/L, 5mmol/ that the mobile phase A, which can be concentration,
L, the ammonium acetate aqueous solution of 5.5mmol/L or 6mmol/L.
In a specific embodiment, mobile phase A is the ammonium acetate aqueous solution of 5mmol/L；Mobile phase B is methanol.
In a preferred embodiment, column size is 3.0mm × 100mm.
In a preferred embodiment, sample volume is 15~25 μ l；Further preferably 18~22 μ l；More preferably 20
μl。
In a preferred embodiment, flow velocity is 0.3~0.7ml/min；Preferably, flow velocity is 0.4~0.6ml/
min；More preferably 0.5ml/min.
In a preferred embodiment, column temperature control is at 39~41 DEG C；More preferably 40 DEG C.
In a specific embodiment, the source parameters setting of Mass Spectrometry Conditions are as follows:
Gas temperature (Gas Temp) is 200 DEG C；Gas flow rate (Gas Flow) is 14l/min；Spray pressure power
It (Nebulizer) is 35psi；Sheath temperature degree (Sheath Gas Heater) is 350 DEG C；Sheath gas (Sheath Gas
It Flow) is 11l/min；Capillary voltage (Caplliary): 3000v；Spray nozzle voltage 1500v.
In a specific embodiment, one of target compound at least following compounds of the method detection or
It is a variety of:
Chlopyrifos (Chlorpyrifos), chlorpyrifos-methyl (Chlorpyrifos-methyl), chlopyrifos
(Chlorpyrifos) and the chloro- 2 hydroxy pyrimidine of metabolin 3,5,6- tri- of chlorpyrifos-methyl (Chlorpyrifos-methyl)
(TCPY), p-nitrophenol (NTP), pentachlorophenol (PCP), bisphenol-A (BPA), Benzophenone -3 (BP-3), triclosan (TCS), right
Methyl hydroxybenzoate (4-OH-methyl ester), ethyl-para-hydroxybenzoate (4-OH-ethyl ester), para hydroxybenzene
Propyl formate (4-OH-propyl ester), phthalic acid diethyl ethyl phosphonate (MEHP), triclocarban (TCC), 3- benzene oxygen
Yl benzoic acid (3-PBA), 3- (2,2- dichloroethylene) -2,2- dinethyl cyclopropane carboxylic acid (DCCA), Methomyl
(Methomyl), specific metabolic object 3- hy droxy furan pellet (3-OH-Carbofuran), Acetamiprid of carbofuran
(Acetamiprid), the major metabolite N- demethyl Acetamiprid (N-DeM-Acetamiprid) of Acetamiprid, ant worm woods
(Imidacloprid), the chloro- 1,3- thiazoline -5- carboxylic acid (2-Cl-1,3-T-5-CA) of 6- chlorine apellagrin (6-CN), 2-.These mesh
The specifying information for marking compound is as shown in table 1.
The classification of 1 22 kinds of target compounds of table
In a specific embodiment, the selection of the internal standard compound is added according to target compound detected.
General requirement is that internal standard compound should be stable and non-sample endogenous material, and retention time should be with mesh detected
It is close to mark the retention time of the compound, optimal is that chemical property is almost the same with target compound.If targeted detected
Close object be it is multiple, and each target compound retention time difference it is larger when, can choose that multiple internal standard compounds are added is right respectively
Should in different reservation periods, when calculating the content of a certain target compound should with retention time and the target compound compared with
For close internal standard compound as a reference to calculating.More preferably, internal standard compound can be the same position of 22 kinds of target compounds in table 1
Plain marker, an internal standard compound can be used for the one or more target compound detected that retention time is closer to therewith
Content calculating.
In a specific embodiment, the internal standard compound is one or more.
In a specific embodiment, the internal standard compound is the isotopic label of target compound detected, is seen
It is in the isotopic label conduct for joined corresponding target compound shown in YES in " Inner stander " in table 3 " item
Mark object.
In a specific embodiment, the content of target compound detected draws standard curve according to internal standard method
It calculates.
In a specific embodiment, 22 kinds of targeteds of various concentration are separately added into urine specimen to be detected
At the mixing mark night for closing object, it is made into the urine specimen of various concentration, it is not interior on the same day continuously to do 5 batches, it investigates 22 kinds of compounds and exists
A certain range of linear dependence calculates the content of target compound detected according to linear equation curve.
One of the major technical difficulty that the exploitation of analysis method of the invention faces are as follows: analysis method will be quick and precisely.By
The detection of a large amount of biological samples in epidemiological study to be applied to, so method efficiency wants high, it may be assumed that pre-treating method wants letter
Just quickly, instrument analysis part will realize that single needle sample introduction in a short time carries out the multiple target compounds object at least table 1
Separate simultaneously accurate quantitative analysis.And urine specimen pre-treatment is often more complicated cumbersome in Existing methods, is needed mostly to urine specimen
After carrying out enzymolysis processing, it is also necessary to which by sequence of operations, Solid Phase Extraction is filtered, is enriched with again, and it is equal that instrument analyzes the single needle time
Long, some even single needle sample introductions reach 30min.
The two of the major technical difficulty that the exploitation of analysis method of the invention faces are as follows: trace sample detects multiple pollutant
Method sensitivity want high.Since all compounds in the biological sample in epidemiological survey in table 1 belong to trace water
It is flat, so method sensitivity wants height to can be only achieved trace pattern detection demand.In view of biological sample involved in epidemiological survey
This sampling is all extremely difficult, and sample size is all fewer, and sample resource is very precious, so as far as possible with minimal amount of pattern detection ratio
More pollutant.Existing methods are differed with urine specimen amount 2ml-4ml, bigger with sample amount, quick with the few method woods of sample amount
It spends also relatively low.
During developing method of the invention, it has been found that the matrix interference in sample is influence method sensitivity
One of principal element, so it is very crucial that matrix, which is removed, during Sample pretreatment.
Existing method generally uses Solid Phase Extraction (SPE) technology, that is, chooses the chromatographic column of certain material filler, to urine sample into
Row filtering, is adsorbed on target compound on small column packing, is then rinsed with particular solution while being adsorbed on the base on filler
Matter finally elutes the target compound being adsorbed on filler with particular solution, machine testing is carried out after concentrate eluant.Due to
22 kinds of object chemical property difference that this method is related to are big, different, are difficult to find and a kind of meet all compound properties
SPE pillar.Even if finding suitable SPE column, not only complicated and time consumption, target compound also lose such method: firstly, urine
Sample can not be adsorbed on completely on small column packing when passing through SPE pillar；Secondly, can also take away one when eluting matrix
Partial target compound.Institute to reach in this way higher sensitivity generally require it is also bigger with sample size.
In order not to lose target compound, the present invention takes urine to be directly centrifuged loading first in method development process,
Testing result finds that the compound responsiveness of compound and the smaller appearance of polarity rearward of the larger appearance of polarity very early is very low, such as schemes
Shown in 1, the peak type and separating degree at each peak are all very poor.The analysis found that urine mesostroma can be a large amount of as organic phase improves
It goes out, enters mass spectrum with target compound to the responsiveness of interfering compound.Mobile phase when being rushed out according to matrix
In organic Phase Proportion it is concluded that these matrix may be a small amount of small molecular protein for containing in urine.
So next we take the operation for carrying out albumen precipitation to urine, three chloroethenes of 2 times of volumes are used first
Acid and perchloric acid do not reach effect, and having carried out dilution to urine sample instead further reduced target compound sensitivity.For
Achieve the purpose that sediment matrix not diluted urine sample again, we are used again with the progress of volatile organic solvent acetonitrile
Albumen precipitation, after 3 times of volumes of acetonitrile and sufficient vortex is added in urine specimen, there is precipitating in centrifugation discovery lower layer, after taking supernatant
It is put into traditional vacuum concentrating instrument and volatilizes acetonitrile, machine testing in remaining urine sample, as a result, it has been found that the appearance in higher organic Phase Proportion
Compound responsiveness has obtained very big improvement, illustrates that the matrix such as corresponding small molecular protein of jamming target compound are in urine
It is removed, and the compound of polarized big appearance morning is responded still without being improved, as shown in Figure 2.
Analysis reason may be that the pH value for the matrix either urine that some polarity are big in urine affects this again
The response of a little compounds, random start debug sample system pH value, finally determine the second containing 1% formic acid by test of many times
Nitrile goes in precipitating urine sample have the matrix of interference effect that can perfectly solve the problems, such as matrix interference, makes the response of each compound
It is attained by detection demand, as shown in Figure 3.
During developing method of the invention, the reasonable set of instrument parameter is also the factor of influence method sensitivity
One of.In order to complete the separation of each target compound within the shorter time as far as possible, we select column to imitate higher C18
Reverse-phase chromatographic column；And mobile phase part by test of many times discovery methanol as organic phase than acetonitrile as organic phase when each chemical combination
Object response is strong.And from the point of view of comprehensive each target compound peak type and responsiveness, the effect when ammonium acetate of 5mmol/L being added in water phase
Most preferably.Since detected target compound parent ion has cation to have anion again, so our mass spectrum parts use EFI
Dynamic polyion detection pattern (DMRM) under mist ion source (ESI), in positive negative mode and use and multiple target compounds simultaneously
Each compound response can be relatively high and more stable under DMRM mode when monitoring.
Method provided by the invention is compared with other methods few with sample size, and pre-treatment is simple and quick, single needle sample analysis
Time is short, and 18min can complete whole appearances, and method high sensitivity (LOD value is relatively low), can save human and material resources into
This, the mass detection of biological sample suitable for epidemiological study.This method is established and is successfully applied to after the completion of examining
The pollutant monitoring of pregnancy period glaucosuria sample in 192 Shanghai City children's aristogenesis queued entries, the result is shown in tables 10.
Method of the invention has the characteristics that versatile, high sensitivity, stability and favorable reproducibility, particularly suitable for height
The analysis detection of throughput sample.
Detailed description of the invention
Fig. 1 is that total ion current (TIC) map of direct loading is centrifuged after urine mark-on, it can be seen that the peak type at each peak in figure
It is all very poor with separating degree.
Fig. 2 is total ion current (TIC) map of urine mark-on loading after acetonitrile treatment, as can be seen from Figure map
Middle section and the later compound peak type and separating degree of back segment appearance are improved, and the compound of polarized big appearance morning does not obtain
Improve.
Fig. 3 is total ion current (TIC) map of loading after the acidified acetonitrile treatment of urine mark-on, as can be seen from Figure
Each compound peak type and separating degree are effectively improved.
Fig. 4 A, 4B, 4C are the matter that method of the invention in embodiment one detects that 22 kinds of target compounds mixing standard liquids obtain
Compose MRM map.
Fig. 5 is that method detection urine specimen of the invention in embodiment two adds 22 kinds of target compounds to mix standard liquid (40ng/
Ml the overlay chart of MRM mass spectrogram after processing is measured after).
Fig. 6 is that method detection water blanks of the invention in embodiment two measure mass spectrogram, and each curve is each mesh in figure
Mark the overlay chart of compound MRM figure.
Specific embodiment
Present inventor gropes to have obtained and a kind of utilizes LC-MS high pass by extensive research and a large amount of experiment
The method of a variety of different type environmental contaminants in amount detection urine, through current experimental verification, this method can be effective for
At least detection and quantitative analysis of 22 kinds of environmental contaminants and/or its body metabolism product (as shown in table 1), it is applied widely,
Versatility is high；Each environmental contaminants and/or the production of its body metabolism all have higher sensitivity, can satisfy trace pattern detection
Demand；The method that each environmental contaminants and/or its body metabolism produce is linearly good, R2>=0.99, quantitative detection result is accurate；
The precision of this method is high, stability is good, is suitable for the long-time sample detection analysis of high-volume sample, be suitable for it is high-throughput into
Sample analysis.
The detection method that the present invention uses, chromatographic condition are as follows:
(1) it Sample pretreatment: takes the urine specimen of certain volume that internal standard compound is added, the vinegar of half urine sample volume is added
Sour ammonium/acetate buffer salt tune ph to 5.0 or so is added β glucuronic acid esterase and is incubated for a period of time；Add after incubation
Entering the acetonitrile of 0.1~2% formic acid containing volume fraction, centrifuging and taking supernatant after sufficient vortex carries out traditional vacuum concentration to supernatant,
The sample introduction sample of machine testing in acquisition；
(2) using sample introduction sample described in LC-MS sample detection:
Detect liquid-phase condition are as follows: chromatographic column uses reverse phase C18 chromatographic column；Mobile phase includes mobile phase A and Mobile phase B；Stream
The formic acid aqueous ammonium that dynamic phase A is 4~6mmol/L；Mobile phase B is methanol；Using gradient elution, gradient elution program is by following
Program carries out: mobile phase A+Mobile phase B=100%；Mobile phase original ratio is 95% mobile phase A and 5% Mobile phase B；0
~4min, Mobile phase B percent by volume are incremented to 10% by 5%；4~12min, Mobile phase B percent by volume are incremented by by 10%
To 60%；12~14min, Mobile phase B percentage by volume are incremented to 85% by 60%；14~16min, Mobile phase B volume basis
Number is incremented to 90% by 85%；16~18min, it is 90% that Mobile phase B, which keeps percentage by volume,；
Detect Mass Spectrometry Conditions are as follows: using the dynamic polyion detection pattern (DMRM) under electric spray ion source (ESI).
It in a specific embodiment, is using ammonium acetate aqueous solution tune pH in step (1).Preferably, the acetic acid
The concentration of aqueous ammonium is 1mol/L.
In a specific embodiment, in step (1), incubation conditions are 37 DEG C of 2~4h of incubation.Preferably, it is incubated for 3h.
In a specific embodiment, in step (1), the addition volume of the acetonitrile of 0.1~2% formic acid is urine specimen
3~8 times of volume.Preferably 4~7 times.More preferably 5~6 times.
In a specific embodiment, in step (1), the volume fraction of formic acid is 0.5~1.5% in the acetonitrile.
Preferably 0.8~1.2%.More preferably 0.9~1.1%.
In a specific embodiment, it is 4mmol/L, 4.5mmol/L, 5mmol/ that the mobile phase A, which can be concentration,
L, the ammonium acetate aqueous solution of 5.5mmol/L or 6mmol/L.
In a specific embodiment, mobile phase A is the ammonium acetate aqueous solution of 5mmol/L；Mobile phase B is methanol.
In a preferred embodiment, column size is 3.0mm × 100mm.
In a preferred embodiment, sample volume is 15~25 μ l；Further preferably 18~22 μ l；More preferably 20
μl。
In a preferred embodiment, flow velocity is 0.3~0.7ml/min；Preferably, flow velocity is 0.4~0.6ml/
min；More preferably 0.5ml/min.
In a preferred embodiment, column temperature control is at 39~41 DEG C；More preferably 40 DEG C.
In a specific embodiment, the source parameters setting of Mass Spectrometry Conditions are as follows: gas temperature (Gas Temp)
It is 200 DEG C；Gas flow rate (Gas Flow) is 14l/min；Spray pressure power (Nebulizer) is 35psi；Sheath temperature degree
(Sheath Gas Heater) is 350 DEG C；Sheath gas (Sheath Gas Flow) is 11l/min；Capillary voltage
(Caplliary): 3000v；Spray nozzle voltage 1500v.
Clear, complete description will be carried out to technical solution of the present invention below, it is clear that described embodiment is this hair
Bright a part of the embodiment, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art exist
Every other embodiment obtained under the premise of creative work is not made, shall fall within the protection scope of the present invention.
Embodiment one
Sample pretreatment step are as follows: after inner mark solution is added in 100ul urine specimen, add the ammonium acetate water of 50ul 1mol/L
Solution (PH is adjusted to 5.0) adds β glucuronic acid esterase, 37 DEG C of incubation 3h after mixing.Incubation finishes taking-up and 500ul is added
Acetonitrile containing 1% formic acid, sufficient vortex 15s, centrifuging and taking supernatant carry out traditional vacuum to supernatant and are concentrated into 100ul, upper machine examination
It surveys, sample volume 20ul.This method is easy to operate, few with sample amount, overcomes that pre-treatment very complicated in other methods is time-consuming and expense
The characteristics of sample size.
Liquid-phase condition: chromatographic column specification: C18 reverse chromatograms column, 3.0mm*100mm；Water phase is the ammonium acetate water of 5mmol/L
Solution, organic phase are pure methanol, and mobile phase original ratio is 95% water phase and 5% organic phase, and when 4min, which is upgraded to 10%, to be had
Machine phase, 12min are upgraded to 60% organic phase, and 14min is upgraded to 85% organic phase, and 16min keeps the ratio after being upgraded to 90% organic phase
2min, rear runing time 4min make mobile phase return to original ratio and reach balance.20 μ L of sample volume.
Mass spectrometry parameters: using the source ESI DMRM ± mode, and design parameter is shown in Table 2.
2 source parameters of table
| Parameter | Value(+) | Value(-) |
| Gas Temp(℃) | 200 | 200 |
| Gas Flow(l/min) | 14 | 14 |
| Nebulizer(psi) | 35 | 35 |
| SheathGasHeater | 350 | 350 |
| SheathGasFlow | 11 | 11 |
| Capillary(V) | 3000 | 3000 |
| Nozzle voltage(v) | 1500 | 1500 |
Method of the invention can detecte 22 kinds of target compounds in urine specimen and carry out quantitative detection and analysis, can be used for
Comprehensive and accurate assessment is carried out to crowd's pollutant.Ion pair information of 22 kinds of target compounds in mass spectrum is as shown in table 3.
3 22 target compound detection ion pairs of table and other parameter informations
The mass spectral results of 22 kinds of target compounds are detected as shown in figs. 4 a-4 c using method of the invention.
Two method specificity of embodiment
Method specificity: blank distilled water and urine mark-on sample are surveyed using method of the invention, see that target compound goes out
Whether peak position has Interference Peaks appearance, to judge this method to the specificity of each target compound.Experimental result such as Fig. 5 and Fig. 6
It is shown.
It is found that method specificity of the invention is good, the peak shape of each target compound is good, uses the party by comparison diagram 5 and Fig. 6
Method measure blank water sample target compound go out peak position do not interfere with peak occur, method high-specificity.
Three matrix effect of embodiment
Matrix effect detection: respectively with the directly upper machine testing of urine specimen mark-on, urine specimen mark-on by before this method
Machine testing on upper machine testing after removing matrix, the certain density sample of distilled water preparation of matrix is handled, is measured respectively each
Target compound response sees influence that matrix responds each target compound and this method to the removal effect of matrix.Inspection
The results are shown in Table 4 for survey.
The matrix effect of 4 22 kinds of target compounds of table
Experimental result is shown: being influenced maximum target compound substrate inhibition by matrix when urine specimen directly goes up machine testing
Rate is up to 92.5%, and response obviously becomes larger after Sample pretreatment mode in this method is handled.Wherein, most of
The response of target compound is improved to original 1.5 times or more, and the response of partial target compound improves to original 3~
4 times or more.This illustrates that matrix effect is greatly improved, and matrix interference becomes smaller, and the sensitivity of compound test is higher.
Example IV LOD value is investigated
LOD value is investigated: the LOD value of 22 kinds of target compounds is investigated using method of the invention.The smaller detection of LOD value is sensitive
It spends higher.Test result is as shown in table 5.
The LOD value of 5 22 kinds of target compounds of table
Experimental result is shown: method of the invention is for when detecting above-mentioned 22 kinds of compounds, LOD value to be in 0.003ng/ml-
Between 0.2ng/ml, wherein the detection sensitivity of each compound is all between the 0.003ng/ml-0.1ng/ml of 21 compounds
It is very high.
Embodiment five is linearly investigated
It is linear to investigate: it is separately added into the mixing mark night of 22 kinds of target compounds of various concentration in human urine sample kind,
It is made into the urine specimen of various concentration.On the same day in continuously do 5 batches, investigate 22 kinds of compounds in a certain range linear
Correlation.Test result is as shown in table 6.
The linear investigation of 6 22 kinds of target compounds of table
| Compound | Mark bent range (ng/ml) | Regression equation (n=5) | R2(n=5) | Weight |
| 2-Cl-1,3-T-5-CA | 0.1-100 | Y=1.07X+4.28*10-3 | 0.997 | 1/x2 |
| 6-CN | 0.1-100 | Y=1.23X+7.06*10-3 | 0.998 | 1/x2 |
| Methomyl | 0.5-100 | Y=0.60X+2.81*10-3 | 0.995 | 1/x2 |
| Imidacloprid | 0.01-100 | Y=0.13X-2.67*10-4 | 0.997 | 1/x2 |
| NTP | 0.01-100 | Y=0.26X-7.34*10-4 | 0.998 | 1/x2 |
| N-DeM-Acetamiprid | 0.1-100 | Y=0.29X+8.75*10-4 | 0.999 | 1/x2 |
| 3-OH-Carbofuran | 0.01-100 | Y=0.20X-5.19*10-4 | 0.995 | 1/x2 |
| Acetamiprid | 0.01-100 | Y=0.63X-1.27*10-4 | 0.996 | 1/x2 |
| TCPY | 0.1-100 | Y=2.33X-1.27*10-3 | 0.993 | 1/x2 |
| 3-PBA | 0.1-100 | Y=0.94X-1.59*10-3 | 0.997 | 1/x2 |
| DCCA | 0.1-100 | Y=0.59X+1.53*10-3 | 0.997 | 1/x2 |
| BPA | 0.1-100 | Y=5.10X+1.35*10-2 | 0.997 | 1/x2 |
| PCP | 0.1-100 | Y=0.46X+1.49*10-3 | 0.995 | 1/x2 |
| MEHP | 0.1-100 | Y=0.94X+4.94*10-2 | 0.997 | 1/x2 |
| BP-3 | 0.1-100 | Y=2.08X-1.03*10-2 | 0.995 | 1/x2 |
| Chlorpyrifos-methyl | 0.5-100 | Y=2.31X-2.98*10-2 | 0.995 | 1/x2 |
| TCC | 0.1-100 | Y=1.11X-2.50*10-3 | 0.999 | 1/x2 |
| TCS | 0.1-100 | Y=0.10X+7.07*10-4 | 0.996 | 1/x2 |
| Chlorpyrifos | 0.1-100 | Y=1.05X-8.43*10-3 | 0.999 | 1/x2 |
| 4-OH-methyl ester | 0.5-500 | Y=0.58X-4.00*10-3 | 0.999 | 1/x2 |
| 4-OH-ethyl ester | 0.01-100 | Y=0.73X-3.88*10-3 | 0.999 | 1/x2 |
| 4-OH-propyl ester | 0.1-100 | Y=0.85X-3.16*10-3 | 0.997 | 1/x2 |
Experimental result is shown: each compound is linear good within the scope of 0.01-500ng/ml, linearly dependent coefficient R2>
0.99, the accuracy rate of method of the invention for the quantitative detection of above-mentioned 22 target compounds is high.
Six day to day precision of embodiment and withinday precision are investigated
Day to day precision and withinday precision: precision is continuously to do the Quality Control sample of 5 batches, each batch between in the daytime
Each 3 groups of low, high (0.4ng/ml, 40ng/ml) sample is made, recovery of standard addition (REC%) and RSD% are counted；Withinday precision is
It does on the same day 5 groups low, high, is spaced 2h between every group, calculates the REC% and RSD% of each Quality Control result.Testing result such as table 7A
With shown in table 7B.
The day to day precision of 22 kinds of target compounds of table 7A
The withinday precision of 22 kinds of target compounds of table 7B
Experimental result is shown: the REC% of day to day precision and withinday precision is between 85%-115%, RSD% <
15%, illustrate that method day to day precision and withinday precision of the invention are good, stability and reproducible.
Seven freeze-thaw stability of embodiment is investigated
Freeze-thaw stability is investigated: in the detection of batch samples, the storage and sampling of sample can be related to freeze thawing, may
Adverse effect is brought to the detection of the compound of stability difference.Method of the invention has investigated the freeze thawing 1 of 22 kinds of target compounds
Secondary stability (each 3 groups of low, high Quality Control), 2 stability of freeze thawing (each 3 groups of low, high Quality Control), calculate each result REC% and
RSD%.Test result is as shown in table 8A and table 8B.Low, high two concentration is respectively 0.4ng/ml, 40ng/ml.
1 stability of freeze thawing of 22 kinds of target compounds of table 8A
2 stability of freeze thawing of 22 kinds of target compounds of table 8B
Testing result is shown: the RSD of low, high two concentration (0.4ng/ml, 40ng/ml) Quality Control is respectively less than 15%, REC%
All between 85%-115%, illustrates freeze thawing 1 time, has good stability for 2 times.Method of the invention is influenced very by sample freeze thawing
It is small, stability and favorable reproducibility.
Eight shelf-stability of embodiment is investigated
Freeze-thaw stability is investigated: in the detection of batch samples, sample to be tested can place for a long time, may be to steady
The detection of the compound of qualitative difference brings adverse effect.37 DEG C of placement 4h stability of method (each 3 groups of low, high Quality Control) of the invention
With urine specimen handle well sample injection disc place 48h stability (each 3 groups of low, high Quality Control), calculate each result REC% and
RSD%.Test result is as shown in table 9A and table 9B.Low, high two concentration is respectively 0.4ng/ml, 40ng/ml.
22 kinds of table 9A, 37 DEG C of target compound placement 4h stability
22 kinds of target compound sample injection disc 48h shelf-stabilities of table 9B
Testing result is shown: the RSD of low, high two concentration (0.4ng/ml, 40ng/ml) Quality Control is respectively less than 15%, REC%
All between 85%-115%, 37 DEG C of the sample sample sample injection disc 48h shelf-stabilities placing 4h stability and handling well are good
It is good.Method of the invention is influenced very little, stability and favorable reproducibility by sample standing time, is suitable for the company of batch samples
Continuous detection.
Embodiment nine
After the completion of method of the invention is established and examined, it is successfully applied in 192 Shanghai City children's aristogenesis queued entries
The pollutant monitoring of pregnancy period glaucosuria sample, the result is shown in tables 10.
The level (ng/ml) of 22 kinds of pollutants in 10 Shanghai City children aristogenesis queued entry pregnancy period glaucosuria of table
Method of the invention is used for the detection of above-mentioned project, shows to detect that fast, high-efficient, flux is high, versatile, clever
The characteristics of sensitivity height, accuracy height, stability and favorable reproducibility.
In conclusion the various embodiments described above and attached drawing are only presently preferred embodiments of the present invention, not to limit this
The protection scope of invention, all within the spirits and principles of the present invention, any modification, equivalent substitution, improvement and etc. done all are answered
It is included within the scope of the present invention.
Claims (10)
1. a kind of method using a variety of different type environmental contaminants in LC-MS/MS high throughput detection urine, feature exist
In, comprising the following steps:
(1) it Sample pretreatment: takes the urine specimen of certain volume that internal standard compound is added, ammonium acetate/acetate buffer salt tune ph is added extremely
5.0 or so, β glucuronic acid esterase is added and is incubated for a period of time；0.1~2% first containing volume fraction is added after incubation
The acetonitrile of acid, centrifuging and taking supernatant after sufficient vortex carry out traditional vacuum concentration to supernatant, the sample introduction sample of machine testing in acquisition
This；
(2) using sample introduction sample described in LC-MS/MS sample detection:
Detect liquid-phase condition are as follows: chromatographic column uses reverse phase C18 chromatographic column；Mobile phase includes mobile phase A and Mobile phase B；Mobile phase A
For the ammonium acetate aqueous solution of 4~6mmol/L；Mobile phase B is methanol；Using gradient elution, gradient elution program presses following procedure
It carries out: mobile phase A+Mobile phase B=100%；Mobile phase original ratio is 95% mobile phase A and 5% Mobile phase B；0~
4min, Mobile phase B percent by volume are incremented to 10% by 5%；4~12min, Mobile phase B percent by volume are incremented to by 10%
60%；12~14min, Mobile phase B percentage by volume are incremented to 85% by 60%；14~16min, Mobile phase B percentage by volume
90% is incremented to by 85%；16~18min, it is 90% that Mobile phase B, which keeps percentage by volume,；
Detect Mass Spectrometry Conditions are as follows: using the dynamic polyion detection pattern under electric spray ion source.
2. the method as described in claim 1, which is characterized in that be molten using ammonium acetate/acetate buffer in the step (1)
Liquid tune pH；Preferably, the concentration of the ammonium acetate/hac buffer is 1mol/L.
3. the method as described in claim 1, which is characterized in that in step (1), incubation conditions are 37 DEG C of 2~4h of incubation；It is preferred that
, it is incubated for 3h.
4. the method as described in claim 1, which is characterized in that in step (1), the addition volume of the acetonitrile of 0.1~2% formic acid
It is 3~8 times of urine specimen volume；Preferably 4~7 times；More preferably 5~6 times.
5. the method as described in claim 1, which is characterized in that in step (1), the volume fraction of formic acid is in the acetonitrile
0.5~1.5%；Preferably 0.8~1.2%；More preferably 0.9~1.1%.
6. the method as described in claim 1, which is characterized in that mobile phase A is the ammonium acetate aqueous solution of 5mmol/L；Mobile phase B
For methanol.
7. the method as described in claim 1, which is characterized in that flow velocity is 0.3~0.7ml/min；Preferably, flow velocity 0.4
~0.6ml/min；More preferably 0.5ml/min.
8. the method as described in claim 1, which is characterized in that column temperature is controlled at 39~41 DEG C；More preferably 40 DEG C.
9. the method as described in claim 1, which is characterized in that the source parameters of Mass Spectrometry Conditions is arranged are as follows:
Gas temperature is 200 DEG C；Gas flow rate is 14l/min；Spray pressure power is 35psi；Sheath temperature degree is 350 DEG C；Sheath gas
Flow velocity is 11l/min；Capillary voltage: 3000v；Spray nozzle voltage 1500v.
10. the method as described in claim 1, which is characterized in that the target compound of the method detection includes at least following
One of compound is a variety of:
Chlopyrifos, chlorpyrifos-methyl, the chloro- 2 hydroxy pyrimidine of 3,5,6- tri-, p-nitrophenol, pentachlorophenol, bisphenol-A, Benzophenone -3,
Triclosan, methyl p-hydroxybenzoate, ethyl-para-hydroxybenzoate, propylparaben, phthalic acid diethyl second
Ester, triclocarban, 3- phenoxy benzoic acid, 3- (2,2- dichloroethylene) -2,2- dinethyl cyclopropane carboxylic acid, Methomyl, 3-
The chloro- 1,3- thiazoline -5- carboxylic acid of hy droxy furan pellet, Acetamiprid, N- demethyl Acetamiprid, ant worm woods, 6- chlorine apellagrin, 2-.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811633115.9A CN109507335B (en) | 2018-12-29 | 2018-12-29 | Method for high-throughput detection of various environmental pollutants in urine by using LC-MS-MS |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811633115.9A CN109507335B (en) | 2018-12-29 | 2018-12-29 | Method for high-throughput detection of various environmental pollutants in urine by using LC-MS-MS |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109507335A true CN109507335A (en) | 2019-03-22 |
| CN109507335B CN109507335B (en) | 2021-11-19 |
Family
ID=65756792
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811633115.9A Active CN109507335B (en) | 2018-12-29 | 2018-12-29 | Method for high-throughput detection of various environmental pollutants in urine by using LC-MS-MS |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109507335B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110514491A (en) * | 2019-08-02 | 2019-11-29 | 暨南大学 | The pre-treating method of organic pollutant in a kind of analysis urine sample |
| CN113009057A (en) * | 2020-10-22 | 2021-06-22 | 重庆医科大学 | Method for detecting neonicotinoid insecticides and metabolites in urine by solid phase extraction-ultra-high performance liquid chromatography-tandem mass spectrometry |
| CN113433227A (en) * | 2021-01-08 | 2021-09-24 | 广州海关技术中心 | Method for detecting 6-chloropicolinic acid in vegetables and fruits |
| 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 |
| CN115078583A (en) * | 2022-06-22 | 2022-09-20 | 江西康美医药保健品有限公司 | Method for detecting content and controlling quality of triclosan in gynecological bacteriostatic lotion |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1192439A1 (en) * | 1999-06-18 | 2002-04-03 | Proteome Systems Ltd. | Apparatus and methods for high resolution separation and analysis of compounds |
| EP1820019A4 (en) * | 2004-11-12 | 2008-05-14 | Pfizer | Method of measuring amyloid-beta peptides |
| CN103901139A (en) * | 2014-04-28 | 2014-07-02 | 中国环境科学研究院 | Pretreatment method for analyzing tetrabromobisphenol A in biologic urine |
| CN106248834A (en) * | 2016-09-23 | 2016-12-21 | 瀚盟测试科技（天津）有限公司 | The UPLC MS/MS detection method of paraben preservative concentration in Urina Hominis |
| CN106680393A (en) * | 2017-01-05 | 2017-05-17 | 上海迪安医学检验所有限公司 | Method for determining contents of 14 types of environmental hormones in urea through liquid chromatography-tandem mass spectrometry |
| CN107389829A (en) * | 2017-09-05 | 2017-11-24 | 环境保护部南京环境科学研究所 | Method and device that is a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos |
-
2018
- 2018-12-29 CN CN201811633115.9A patent/CN109507335B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1192439A1 (en) * | 1999-06-18 | 2002-04-03 | Proteome Systems Ltd. | Apparatus and methods for high resolution separation and analysis of compounds |
| EP1820019A4 (en) * | 2004-11-12 | 2008-05-14 | Pfizer | Method of measuring amyloid-beta peptides |
| CN103901139A (en) * | 2014-04-28 | 2014-07-02 | 中国环境科学研究院 | Pretreatment method for analyzing tetrabromobisphenol A in biologic urine |
| CN106248834A (en) * | 2016-09-23 | 2016-12-21 | 瀚盟测试科技（天津）有限公司 | The UPLC MS/MS detection method of paraben preservative concentration in Urina Hominis |
| CN106680393A (en) * | 2017-01-05 | 2017-05-17 | 上海迪安医学检验所有限公司 | Method for determining contents of 14 types of environmental hormones in urea through liquid chromatography-tandem mass spectrometry |
| CN107389829A (en) * | 2017-09-05 | 2017-11-24 | 环境保护部南京环境科学研究所 | Method and device that is a kind of while detecting 3 kinds of metabolites of urine Chlorpyrifos |
Non-Patent Citations (5)
| Title |
|---|
| ALEXANDROS G. ASIMAKOPOULOS ET AL: "A multiclass bioanalytical methodology for the determination of bisphenol A diglycidyl ethers, phydroxybenzoic acid esters, benzophenonetype ultraviolet filters, triclosan, and triclocarban in human urine by liquid chromatographytandem mass spectrometry", 《JOURNAL OF CHROMATOGRAPHY A》 * |
| LINDA SCHLITTENBAUER ET AL: "Ultrasound-assisted hydrolysis of conjugated parabens in human urine and their determination by UPLC–MS/MS and UPLC–HRMS", 《ANAL BIOANAL CHEM》 * |
| NANCY K. WILSON ET AL: "Exposures of preschool children to chlorpyrifos, diazinon, pentachlorophenol, and 2,4-dichlorophenoxyacetic acid over 3 years from 2003 to 2005: A longitudinal model", 《JOURNAL OF EXPOSURE SCIENCE AND ENVIRONMENTAL EPIDEMIOLOGY》 * |
| 张圣虎 等: "高效液相色谱-串联质谱法测定儿童和成人尿液中双酚A、四溴双酚A和辛基酚", 《分析化学 研究报告》 * |
| 赵莎莎 等: "HPLC-MS/MS测定尿液BPA、TCS 和4-n-NP方法的建立及其应用", 《环境与职业医学》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110514491A (en) * | 2019-08-02 | 2019-11-29 | 暨南大学 | The pre-treating method of organic pollutant in a kind of analysis urine sample |
| CN113009057A (en) * | 2020-10-22 | 2021-06-22 | 重庆医科大学 | Method for detecting neonicotinoid insecticides and metabolites in urine by solid phase extraction-ultra-high performance liquid chromatography-tandem mass spectrometry |
| CN113009057B (en) * | 2020-10-22 | 2023-11-07 | 重庆医科大学 | Method for detecting neonicotinoid insecticides and metabolites in urine by solid phase extraction-ultra-high performance liquid chromatography-tandem mass spectrometry |
| CN113433227A (en) * | 2021-01-08 | 2021-09-24 | 广州海关技术中心 | Method for detecting 6-chloropicolinic acid in vegetables and fruits |
| 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 |
| CN115078583A (en) * | 2022-06-22 | 2022-09-20 | 江西康美医药保健品有限公司 | Method for detecting content and controlling quality of triclosan in gynecological bacteriostatic lotion |
| CN115078583B (en) * | 2022-06-22 | 2024-06-07 | 江西康美医药保健品有限公司 | Method for detecting triclosan content and controlling quality in gynecological antibacterial lotion |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109507335B (en) | 2021-11-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109507335A (en) | Utilize the method for a variety of different type environmental contaminants in LC-MS-MS high throughput detection urine | |
| Ramautar et al. | CE–MS for metabolomics: developments and applications in the period 2014–2016 | |
| CN107543877A (en) | A kind of method that SPE liquid chromatography tandem mass spectrometry determines six 24 kinds of antibiotic of class in water body simultaneously | |
| CN109060983A (en) | A kind of method of liquid chromatography-tandem mass spectrometry detection metanephrine substance | |
| Seiler | Assay procedures for polyamines in urine, serum, and cerebrospinal fluid. | |
| Şentürk et al. | Analytical methods for determination of selective serotonin reuptake inhibitor antidepressants | |
| CN113176364A (en) | Method for simultaneously detecting trimethylamine oxide and phenylacetylglutamine, detection kit and application thereof | |
| CN110652749B (en) | Composite nanofiber online micro solid-phase extraction column and preparation method thereof | |
| Zhang et al. | In vivo monitoring of the monoamine neurotransmitters in rat brain using microdialysis sampling with liquid chromatography electrochemical detection | |
| CN113533549A (en) | White spirit taste substance identification and analysis system | |
| CN109254053B (en) | Preparation method and application of environmental estrogen electrochemical analysis sensor | |
| Rosting et al. | High-throughput analysis of drugs in biological fluids by desorption electrospray ionization mass spectrometry coupled with thin liquid membrane extraction | |
| CN109254052B (en) | Preparation method and application of electrochemical luminescence sensor for organophosphorus pesticide | |
| Jaenicke et al. | Determination of lead in blood by hydrodynamic voltammetry in a flow injection system with wall-jet detector | |
| Xie et al. | Comparison of solid-phase microextraction and liquid–liquid extraction in 96-well format for the determination of a drug compound in human plasma by liquid chromatography with tandem mass spectrometric detection | |
| CN105334282B (en) | Co-detecting method for environmental estrogens in surface water body | |
| Ji et al. | A thin-layer electrochemical detector coated with nafion film for liquid chromatography | |
| CN112946119A (en) | Method for analyzing and detecting 11 phenoxy carboxylic acid herbicides in environmental water sample | |
| Li et al. | Determination of trimethylamine in fish by capillary electrophoresis with electrogenerated tris (2, 2′‐bipyridyl) ruthenium (II) chemiluminescence detection | |
| CN109254058B (en) | Preparation method and application of organophosphorus pesticide sensor based on nickel nitride array | |
| CN112067685B (en) | Method for rapidly detecting clenbuterol in meat by Fapex-TD-ESI-MS/MS | |
| CN110702494B (en) | Preparation method and application of triphenylmethane dye and high-selectivity solid-phase microextraction probe of metabolic product of triphenylmethane dye | |
| Wang et al. | A novel and sensitive screening method for β-agonists in porcine urine by using atmospheric solid analysis probe source coupled tandem mass spectrometry | |
| Furuumi et al. | Ionspray ionization–mass spectrometric separation and determination of heptafluorobutyryl derivatives of polyamines | |
| CN112697898B (en) | Method for rapidly determining content of ustilagin A in urine or cell culture medium by liquid chromatography-mass spectrometry |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |