CN109342624A - The method that Solid Phase Extraction pre-treatment combination LC-MS technology detects 15 kinds of antibiotic in aquaculture system simultaneously - Google Patents
The method that Solid Phase Extraction pre-treatment combination LC-MS technology detects 15 kinds of antibiotic in aquaculture system simultaneously Download PDFInfo
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Abstract
The present invention provides a kind of methods that Solid Phase Extraction pre-treatment combination LC-MS technology detects 15 kinds of antibiotic in aquaculture system simultaneously.This method comprises: (1) water sample pre-processes, sulfamethoxazole-is added13C6, Norfloxacin-D5, potassium penicillin G-D5, tetracycline-D6, olaquindox-D4, Amoxicillin-13C6With trimethoprim-D3The internal standard compound of this seven kinds instruction rate of recovery；(2) solid-phase extraction column enrichment purification target antibiotic is used；(3) content of seven 15 kinds of antibiotic of class in LC-MS instrument detection aquaculture water is used using internal standard method.This method uses experiment of single factor, pass through preferred internal standard compound, optimizing detection condition, can disposably simultaneously detect in aquaculture system containing eight kinds of newly-increased antibiotic 15 kinds of antibiotic residual condition, have many advantages, such as high precision, high sensitivity, high stability, it is highly selective it is low with detection limit, testing result is more true and reliable.
Description
Technical field
The invention belongs to the detection technique field of water environment pollution object, it is related in a kind of aquaculture system while detects trace
It measures the detection method of 15 kinds of common antibiotics more particularly to a kind of Solid Phase Extraction pre-treatment combination LC-MS technology while detecting
The method of 15 kinds of antibiotic in aquaculture system.
Background technique
The 1950s is started from using antibiotic treatment and prevention bacterial disease in aquaculture.With aquaculture
The rapid development of industry, the occurrence frequency of disease is higher and higher in breeding process, use scope and dosage of the antibiotic as veterinary drug
Increasingly increase.Increase with culturists to the economic interests pursuit and aquatile drug resistance of aquaculture, antibiotic
Dosage is continuously improved, or even is used with unreasonable high dose.
The adverse effect of abuse of antibiotics and increasingly severe trend is shown to the harm of the mankind, and is increasingly becoming people
One hot spot of society of common concern.According to statistics, about 750-1000t aureomycin, 500-700t soil are mould every year in China
Element, 400t Norfloxacin, 100t Ciprofloxacin, 15t Ofloxacin are used for the feed addictive of animal.Existing research shows
2003, the total output of China's sulfa antibiotics broke through 2000t, wherein about 4500t makes as veterinary drug within Chinese territory
With.As can be seen that the usage quantity of antibiotics is quite surprising in current aquaculture industry of China.In highdensity aquaculture
In the industry, antibiotic 70-80% used in aquaculture process eventually enters into water environment.
As the mankind are for the pay attention to day by day of water environment safety problem, in relation in environment water antibiotics leftover detection it is special
Benefit and paper, which are disclosed, to be delivered, but these research spininess to certain a kind of or two three classes antibiotic Synchronization Analysis and are analyzed
The total quantity of antibiotic is most few.However the number of species of antibiotic are various, point about multiple types multi-quantity antibiotic residue
Seem rather important from enrichment and the foundation of synchronized analyzing method.
High performance liquid chromatography tandem mass spectrometry can be according to characteristic ion in quantitative analysis and thus characteristic ion is broken
Fingerprint of the fragments characteristic of generation as a certain target compound, it is screened from the blend sample of complex mechanism into
Row is quantitative, ensure that the accuracy and precision of detection.
As antibiotic largely uses in aquaculture process, for can quick, sensitive, accurately measure various water
Produce antibiotic residue as much as possible in breeding water body exist there is an urgent need to.However, at present for a variety of in aquaculture system
The separation and concentration of antibiotic and altogether detection there is no the reliable detection method of complete set, the existing generally existing shakiness of detection method
Determine, interfere the problems such as more, detection antibiotic value volume and range of product is limited, the rate of recovery is limited, therefore, it is steady to be badly in need of a kind of testing result
It is fixed, interfere it is small, can be quick, efficient, sensitive, accurate, while detecting that antibiotic as much as possible is residual in aquaculture system
What is stayed includes separation and concentration and the complete reliable detection method that detects altogether.
Summary of the invention
The purpose of the present invention is intended to: overcome it is of the existing technology it is unstable, interference is more, detects antibiotic quantity and recycling
The deficiency for the problems such as rate is limited, provides that a kind of testing result is stable, interference is small, fast and efficiently while detecting aquaculture water
The method of remaining 15 kinds of body kind typical antibiotic, i.e., a kind of Solid Phase Extraction pre-treatment combination LC-MS technology detect water simultaneously
The method for producing 15 kinds of antibiotic in breeding water body.
Technical concept of the invention is as follows:
Method of the invention uses Solid Phase Extraction (SPE) pre-treatment combination high performance liquid chromatography tandem mass spectrum technology (SPE-
HPLC-MS/MS), by the optimization of the optimization of pretreatment process, liquid phase chromatogram condition and Mass Spectrometry Conditions, it finally can be realized standard
Really, stablize, interfere it is small, quickly and efficiently to 15 kinds in aquaculture water sample typical antibiotic while enrichment and quantitative survey
It is fixed.This method uses experiment of single factor, screens suitable solid-phase extraction column, and to activation method during solid phase extraction concentration,
ELUTION METHOD, elution process are optimized, and can disposably detect the residual condition of 15 kinds of antibiotic simultaneously, have high-accuracy
Degree, high sensitivity, high stability, the highly selective advantages such as low with detection limit.The internal standard compound for indicating the rate of recovery is added in by this method
Before Solid Phase Extraction, the loss of target substance in pretreatment process can be indicated well, keeps final result true and reliable.This method
Can be remaining antibiotic in aquaculture system, including sulphadiazine, sulfamethyldiazine, sulphadimethoxine,
Trimethoprim, olaquindox, sulfamethoxazole, Amoxicillin, furazolidone, ciprofloxacin hydrochloride, Norfloxacin, En Nuosha
15 kinds of star, aureomycin, terramycin and Doxycycline Hyclate antibiotic, provide effective accurate detection method.
The purpose of the present invention is achieved through the following technical solutions:
A kind of Solid Phase Extraction pre-treatment combination LC-MS technology of the present invention, which is detected simultaneously in aquaculture system, to be resisted for 15 kinds
The method of raw element, this method comprises: the pretreatment of (1) water sample, is added internal standard compound before object is enriched with and indicates the rate of recovery；(2)
Use solid-phase extraction column enrichment purification target antibiotic；(3) internal standard method is used, LC-MS instrument quantitative detection aquaculture is used
The content of 15 kinds of antibiotic in water body.
A kind of Solid Phase Extraction pre-treatment combination LC-MS technology of the present invention, which is detected simultaneously in aquaculture system, to be resisted for 15 kinds
The method of raw element, specifically carries out as follows:
(1) pretreatment of water sample
Water sample is separately added into sulfamethoxazole-after membrane filtration removes suspended matter13C6, trimethoprim-D3, benzyl penicillin
Sylvite-D5, Norfloxacin-D5, tetracycline-D6, olaquindox-D4, Amoxicillin-13C6The internal standard compound of this seven kinds instruction rate of recovery；
Adjust water sample pH；
(2) solid phase extraction concentration target antibiotic:
Successively use methyl tertiary butyl ether(MTBE), methanol, ultrapure water activated solid extraction column；By processed water sample in step (1)
After the solid-phase extraction column activated；Solid-phase extraction column is eluted with leacheate after the completion of enrichment and is dried in vacuo；Certain volume is used again
Eluting solvent elute object, collect eluent simultaneously dry up under nitrogen flowing, residue obtained to re-dissolve, constant volume；After filtering
It is transferred in brown sample injection bottle, it is to be measured；
(3) high performance liquid chromatography tandem mass spectrometry measures the content of seven 15 kinds of antibiotic of class in water body
By sample to be tested in step (2), using internal standard method, on high performance liquid chromatography tandem mass spectrum instrument, quantitative detection palm fibre
The content for 15 kinds of antibiotic being enriched in color sample injection bottle from water body；15 kinds of antibiotic include aquaculture it is common or by
The substance of disabling, including sulfamido, quinolones, Tetracyclines, beta-lactam, itrofurans, trimethoprim, olaquindox
These seventh types of antibiotic；15 kinds of antibiotic are respectively sulphadiazine, sulfamethyldiazine, sulphadimethoxine, methoxy benzyl
Pyridine, olaquindox, sulfamethoxazole, sulfadimidine, Amoxicillin, furazolidone, ciprofloxacin hydrochloride, Norfloxacin,
Enrofloxacin, aureomycin, terramycin and Doxycycline Hyclate.
Further, in step (1), the sulfamethoxazole-of 100ng/L is separately added into before Solid Phase Extraction13C6, methoxy benzyl
Pyridine-D3, potassium penicillin G-D5, Norfloxacin-D5, tetracycline-D6, olaquindox-D4With Amoxicillin-13C6This seven kinds of internal standard compounds
Indicate the rate of recovery.
Further, in step (1), water sample is first added and refers to after glass fiber filter is filtered to remove graininess suspended matter
Show the internal standard compound of the rate of recovery, then adjust pH value to 3 or so with dilute hydrochloric acid, pH value approximate range is 3 ± 0.1.
Further, in step (1), sample filtering uses aperture for 0.7 μm of Whatman GF/F Series glass fiber
Filter membrane.The purpose is to filter removal particle suspensions matter, prevent from influencing subsequent enrichment and detection to object.
Further, in above-mentioned steps (2), solid-phase extraction column used in enrichment method sample is Waters company Oasis
HLB pillar, adsorbent be by two kinds of monomers of lipophilicity divinylbenzene and hydrophily n-vinyl pyrrolidone by a certain percentage
The macroporous copolymer aggregated into is the universal adsorbent for acidic, neutral and basic compounds.
Further, in above-mentioned steps (2), methyl tertiary butyl ether(MTBE) used in activated solid extraction column, first alcohol and water are used
Amount is respectively 1 times of extraction column volume, and the flow control of activated solid extraction column is in 1-4mL/min；Water sample crosses column flow rate (on water sample
Sample speed) it controls in 3-5mL/min；Pure water used in elution extraction column and low concentration methanol aqueous solution are respectively extraction column volume
1-2 times；Draining after elution, time 2min；Eluting eluting solvent used in solid-phase extraction column is methanol, methanol eluting solvent
Amount be to extract 2 times of column volume, eluent flow rate is controlled in 4-5mL/min.
Further, in above-mentioned steps (2), eluting low concentration methanol used is methanol-of the volumetric concentration less than 5%
Aqueous solution, it is therefore an objective to remove impurity, reduce noise jamming.
Further, it in above-mentioned steps (2), elutes methanol used in solid-phase extraction column and is eluted using HPLC grades of methanol,
After vacuum methanol remaining in pillar is collected.After elution, eluent nitrogen under the conditions of 40 DEG C of heating water bath is blown to close dry.
Further, in above-mentioned steps (2), use volumetric concentration for 70% methanol-water solution constant volume, ultrasound
To be measured in brown sample injection bottle with being transferred to after the filtering of pin type filter after 2min, it is 0.22 μm of polytetrafluoro that pin type filter, which selects aperture,
Ethylene pin type filter.
Further, in above-mentioned steps (3), the Waters of Waters, US is selectedTQ-Smicro high
Effect liquid phase chromatogram tandem mass spectrometer detects 15 kinds of target antibiotic；The chromatographic column of liquid chromatogram selects ACQUITYBEH C18 model column 2.1 × 100mm, 1.7 μm.
Further, in above-mentioned steps (3), liquid chromatogram separation parameter are as follows: 35 DEG C of chromatographic column column temperature；2 μ L of sample volume；Into
10 DEG C of specimen chamber temperature；Flow rate of mobile phase 0.25mLmin-1；Mobile phase A is formic acid-aqueous solution that volumetric concentration is 0.5%,
In water be Milli-Q ultrapure water；Mobile phase B is acetonitrile；Gradient elution program: 0-2.2min, 16%B；2.2-2.5min
16%B-95%B；2.5-5.5min, 95%B；5.5-6.0min, 95%B-16%B；6.0-10.0min, 16%B.
Further, in above-mentioned steps (3), Mass Spectrometer Method condition are as follows: use electric spray ion source positive ion mode ESI
+, triple level four bars mass analyzers, scanning mode: multiple-reaction monitoring pattern MRM is detected；Collision gas is high-purity argon gas；It is de-
Solvent temperature degree is 500 DEG C, and Desolvention gas velocity degree is 1000L/Hr, and orifice potential 30V, capillary voltage 3.5kV are swept
Retouching the time is 0.1S.
Beneficial effects of the present invention are as follows:
Solid Phase Extraction pre-treatment combination high performance liquid chromatography tandem mass spectrum technology (SPE- is used the present invention provides a kind of
HPLC-MS/MS the method for remaining 15 kinds typical antibiotic in aquaculture system) is detected simultaneously.This method is closed by screening
Suitable solid-phase extraction column, and activation method, ELUTION METHOD, elution process during solid phase extraction concentration are optimized, most
It can be realized that high sensitivity, selectivity is good, accuracy is high eventually, this method is suitble to residual antibiotic in practical aquaculture system
Measurement.
The method have the advantages that:
(1) method choice Waters company Oasis HLB solid-phase extraction column of the invention is enriched with 15 kinds of antibiotic
Purification, eliminates interference impurity, reduces matrix effect, improve selectivity and enrichment times；The rate of recovery is high and stablizes, and selects
Property is strong, and adsorption capacity is big, has achieved the purpose that efficiently separate enrichment to target antibiotic.
(2) present invention uses inner mark method ration, and the concentration for measuring antibiotic is more acurrate, and linear relationship is good, and relative standard is inclined
Difference is small, improves the precision of detection and analysis.
It (3), can be very before internal standard substance (instruction rate of recovery substitute) is added in solid phase extraction procedure by method of the invention
The loss of target substance in pretreatment process is indicated well, meanwhile, consider that different classes of antibiotic structure and property exist very
Big difference has different degrees of loss in pretreatment process, so, sulfamethoxazole-is selected respectively13C6As sulfamido
Antibiotic (sulphadiazine, sulfamethyldiazine, sulfamethazine, sulphadimethoxine, sulfamethoxazole)
Internal standard compound selects Norfloxacin-D5As quinolone antibiotics (ciprofloxacin hydrochloride, Norfloxacin, Enrofloxacin)
Internal standard compound selects tetracycline-D6As the internal standard compound of tetracycline antibiotics (aureomycin, terramycin, Doxycycline Hyclate), choosing
With potassium penicillin G-D5As the internal standard compound of Nitrofuran antibiotics (furazolidone), trimethoprim-D is selected3As methoxy
The internal standard compound of benzyl pyridine selects olaquindox-D4As the internal standard compound of olaquindox, Amoxicillin-is selected13C6It is anti-as beta-lactam
The internal standard compound of raw element (Amoxicillin), since internal standard compound is selected appropriately, so that final detection result is more true and reliable.
And in existing organic pollution materials residual minim antibiotic detection method, Tetracyclines and beta-lactam antibiosis
There is no corresponding internal standard substances, usually selection Norfloxacin-D for element5As internal standard substance, the result detected in this way is had
Some deviations.In existing micro antibiotic detection method, the detection of olaquindox, usually using antifebrin as internal standard substance,
Its testing result is not so good as in the present invention with olaquindox-D4The testing result of internal standard compound as olaquindox is accurate.
(4) detection method of the invention, primary upper machine sample introduction can detect a variety of anti-in aquaculture system simultaneously
Raw element (15 kinds of antibiotic), detection process time-consuming is few (time-consuming 10min), and the accuracy and high sensitivity of detection, detection limit is low, can
To detect the residual condition of traces of antibiotic in environment water；Meanwhile the method for the present invention also has higher recovery, it is opposite to mark
Quasi- deviation is smaller, this illustrates the pre-treating method of the application and detection method is reliable and repeating effect is good.
(5) detection method of the invention is easy to operate, and organic reagent usage amount is few, and environmental toxicity is low.
(6) detection method of the invention detects in water body simultaneously in existing Solid Phase Extraction-liquid chromatography tandem mass spectrometry
On the basis of the detection method of multiple types antibiotic, for the 15 kinds of antibiotic commonly used or disabled in aquaculture system as mesh
Mark (contained in this 15 kinds of antibiotic newly-increased sulfamethyldiazine, trimethoprim, olaquindox, furazolidone, Norfloxacin,
Aureomycin, terramycin and Doxycycline Hyclate this eight kinds of antibiotic), which passes through preferred internal standard compound, and optimization inspection
Survey condition disposably can detect to contain 15 kinds of antibiosis including eight kinds of newly-increased antibiotic in aquaculture system simultaneously
Element, also, the detection method internal standard compound rate of recovery is higher, and data result is more true and reliable.
Detailed description of the invention
Fig. 1 is the recovery of standard addition that 15 kinds of antibiotic in water body are measured using three kinds of different eluting solvents.
Specific embodiment
For a better understanding of the present invention, below with reference to the embodiment content that the present invention is furture elucidated, but it is of the invention
Content is not limited solely to the following examples.
Embodiment 1: Solid Phase Extraction eluting solvent selection
Using internal standard method, washing in the detection method of Solid Phase Extraction pre-treatment combination LC-MS technology of the invention is determined
Desolventizing measures three kinds of different eluting solvents to the recovery of standard addition of 15 kinds of antibiotic in aquaculture system.Recovery of standard addition
Experiment is using tap water as sample: adding antibiotic mixed standard solution (antibiotic hybrid standard of 10 μ L into sample respectively
Solution is the mixed standard solution of 15 kinds of antibiotic, wherein the concentration of 15 kinds of antibiotic is respectively 10mg/L).
Solid Phase Extraction pre-treatment combination LC-MS technology of the invention detects 15 kinds of antibiosis in aquaculture system simultaneously
The method of element is specifically implemented according to the following steps:
Step 1: water sample pre-processes
Set up three groups of experiments.Every group of sample sets blank sample and mark-on sample, and sample sets three in parallel.It is accurate respectively to measure 1L water
Sample is in clean beaker.Mark-on sample is separately added into antibiotic hybrid standard liquid by above-mentioned additive amount, i.e., adds respectively into water sample
Add the antibiotic hybrid standard liquid that the concentration of 10 μ L is 10mg/L, makes the ultimate density for the antibiotic being added in water body
100ng/L.Separately set blank group (blank group does not add any antibiotic).
Before Solid Phase Extraction, the sulfamethoxazole-of 100ng is added into sample respectively13C6, trimethoprim-D3, benzyl penicillin
Sylvite-D5, Norfloxacin-D5, tetracycline-D6, olaquindox-D4With Amoxicillin-13C6The internal standard compound of this seven kinds instruction rate of recovery,
It is sufficiently mixed uniformly with water sample.That is: sulfamethoxazole-is selected respectively13C6As between sulphadiazine, sulfamethyldiazine, sulfanilamide (SN)
Dimethoxypyridin, olaquindox, sulfamethoxazole and sulfadimidine internal standard compound, select trimethoprim-D3As methoxy
The internal standard compound of benzyl pyridine selects potassium penicillin G-D5As the internal standard compound of furazolidone, Norfloxacin-D is selected5As cyclopropyl sand
The internal standard compound of star hydrochloride, Norfloxacin and Enrofloxacin selects tetracycline-D6It is how western as aureomycin, terramycin and hydrochloric acid
The internal standard compound of ring element selects olaquindox-D4As the internal standard compound of olaquindox, Amoxicillin-is selected13C6It is anti-as beta-lactam
The internal standard compound of raw element (Amoxicillin).
Sample is sufficiently mixed uniformly after mark-on, is filtered to remove graininess suspended matter through glass fiber filter paper, then with dilute salt
Acid for adjusting pH is to 3 or so.
Step 2: solid phase extraction concentration target antibiotic
Following solid phase extraction procedure is completed using Full-automatic solid phase extraction instrument, and solid-phase extraction column used is Waters company
Oasis HLB pillar, adsorbent are by two kinds of monomers of lipophilicity divinylbenzene and hydrophily n-vinyl pyrrolidone by one
The macroporous copolymer that certainty ratio aggregates into.Solid phase extraction procedure is as follows:
Successively pillar, flow velocity 4mL/min are extracted with 5mL methyl tertiary butyl ether(MTBE), methanol, ultrapure water activated solid.Pre- place
1000mL water sample after reason crosses column with the speed of 5mL/min, after end of the sample, successively uses 6mL ultrapure water and isometric volume
The methanol-water solution that concentration is 5% elutes extraction column, vacuum drying with the speed of 4mL/min.Finally use the eluting solvent of 10mL
Elution, eluting solvent are respectively acetonitrile, methanol and ethyl acetate (being HPLC grades), and flow control is received in 4mL/min, eluent
Into tool plug glass centrifuge tube, eluent (40 DEG C of heating water bath) under water bath condition is dried up under nitrogen stream effect, is obtained collection
Residue is settled to 1mL, ultrasonic 2min with the methanol-water solution that volumetric concentration is 70%, filters through 0.22 μm of PTFE pin type filter
After be transferred in brown sample injection bottle, it is to be measured.
Step 3: high performance liquid chromatography tandem mass spectrometry measures the content of 15 kinds of antibiotic in water body
Using internal standard method, in liquid chromatography-tandem mass spectrometry instrument, 15 kinds of antibiosis in the water sample in quantitative detection sample injection bottle
The concentration of element.
Select the Waters of Waters, USTQ-Smicro high performance liquid chromatography tandem mass spectrum instrument is to 15
Kind target antibiotic is detected；The chromatographic column of liquid chromatogram selects ACQUITYBEH C18 model column 2.1 ×
100mm, 1.7 μm.
Liquid chromatogram separation parameter are as follows: 35 DEG C of chromatographic column column temperature；2 μ L of sample volume；10 DEG C of sample introduction room temperature；Flow rate of mobile phase
0.25mL·min-1；Mobile phase A is formic acid-aqueous solution that volumetric concentration is 0.5%, and water therein is Milli-Q ultrapure water；
Mobile phase B is acetonitrile；Gradient elution program: 0-2.2min, 16%B；2.2-2.5min, 16%B-95%B；2.5-5.5min
95%B；5.5-6.0min, 95%B-16%B；6.0-10.0min, 16%B.
Mass Spectrometer Method condition are as follows: use electric spray ion source positive ion mode ESI+, triple level four bars mass analyzers are swept
Retouch mode: multiple-reaction monitoring pattern MRM is detected；Collision gas is high-purity argon gas；Desolvation temperature is 500 DEG C, desolventizing
Air velocity is 1000L/Hr, orifice potential 30V, capillary voltage 3.5kV, sweep time 0.1S.
Experiment, which is measured, sees Fig. 1 using the rate of recovery of three kinds of different eluting solvents (acetonitrile, methanol and ethyl acetate), from Fig. 1
It can be seen that the rate of recovery for using methanol as eluting solvent in Solid Phase Extraction step is best.Therefore, in aquaculture system antibiotic
In actually detected, methanol can be selected as the eluting solvent in Solid Phase Extraction step.
Embodiment 2: recovery of standard addition experiment
Using internal standard method, inspection is measured by the detection method of Solid Phase Extraction pre-treatment combination LC-MS technology of the invention
Aquaculture system in 15 kinds of antibiotic recovery of standard addition.Recovery of standard addition is tested using tap water as sample: respectively to sample
10 μ L are added in product, (the antibiotic mixed standard solution is the mixing of 15 kinds of antibiotic to the antibiotic mixed standard solution of 20 μ L
Standard solution, wherein the concentration of 15 kinds of antibiotic is respectively 10mg/L).
Solid Phase Extraction pre-treatment combination LC-MS technology of the invention detects 15 kinds of antibiosis in aquaculture system simultaneously
The method of element is specifically implemented according to the following steps:
Step 1: water sample pre-processes
Set up three kinds of spiked levels (three groups of samples).Every group of sample sets two in parallel, and the accurate 1L water sample that measures is in clean respectively
In net beaker.Wherein two groups of samples are separately added into antibiotic hybrid standard liquid by above-mentioned additive amount, i.e., add respectively into water sample
10 μ L, 20 μ L concentration be the antibiotic hybrid standard liquid of 10mg/L, make the ultimate density point for the antibiotic being added in water body
It Wei not 100ng/L and 200ng/L.Third group sample is set as blank group (blank group does not add any antibiotic).
Before Solid Phase Extraction, the sulfamethoxazole-of 100ng is added into sample respectively13C6, trimethoprim-D3, benzyl penicillin
Sylvite-D5, Norfloxacin-D5, tetracycline-D6, olaquindox-D4With Amoxicillin-13C6The internal standard compound of this seven kinds instruction rate of recovery,
It is sufficiently mixed uniformly with water sample.That is: sulfamethoxazole-is selected respectively13C6As between sulphadiazine, sulfamethyldiazine, sulfanilamide (SN)
Dimethoxypyridin, olaquindox, sulfamethoxazole and sulfadimidine internal standard compound, select trimethoprim-D3As methoxy
The internal standard compound of benzyl pyridine selects potassium penicillin G-D5As the internal standard compound of furazolidone, Norfloxacin-D is selected5As cyclopropyl sand
The internal standard compound of star hydrochloride, Norfloxacin and Enrofloxacin selects tetracycline-D6It is how western as aureomycin, terramycin and hydrochloric acid
The internal standard compound of ring element selects olaquindox-D4As the internal standard compound of olaquindox, Amoxicillin-is selected13C6It is anti-as beta-lactam
The internal standard compound of raw element (Amoxicillin).
Sample is sufficiently mixed uniformly after mark-on, is filtered to remove graininess suspended matter through glass fiber filter paper, then with dilute salt
Acid for adjusting pH is to 3 or so.
Step 2: solid phase extraction concentration target antibiotic
Following solid phase extraction procedure is completed using Full-automatic solid phase extraction instrument, and solid-phase extraction column used is Waters company
Oasis HLB pillar, adsorbent are by two kinds of monomers of lipophilicity divinylbenzene and hydrophily n-vinyl pyrrolidone by one
The macroporous copolymer that certainty ratio aggregates into.Solid phase extraction procedure is as follows:
Successively pillar, flow velocity 4mL/min are extracted with 5mL methyl tertiary butyl ether(MTBE), methanol, ultrapure water activated solid.Pre- place
1000mL water sample after reason crosses column with the speed of 5mL/min, successively dense with 6mL ultrapure water and isometric volume after end of the sample
Degree elutes extraction column, vacuum drying for 5% methanol-water solution with the speed of 4mL/min.Finally use the methanol (HPLC of 10mL
Grade methanol) elution, flow control is in 4mL/min, and eluent is collected into tool plug glass centrifuge tube, and eluent is under water bath condition
(40 DEG C of heating water bath) dries up under nitrogen stream effect, and obtained residue is settled to the methanol-water solution that volumetric concentration is 70%
1mL, ultrasonic 2min are transferred in brown sample injection bottle after 0.22 μm of PTFE pin type filter filters, to be measured.
Step 3: high performance liquid chromatography tandem mass spectrometry measures the content of 15 kinds of antibiotic in water body
Using internal standard method, in liquid chromatography-tandem mass spectrometry instrument, 15 kinds of antibiosis in the water sample in quantitative detection sample injection bottle
The concentration of element.
Select the Waters of Waters, USTQ-Smicro high performance liquid chromatography tandem mass spectrum instrument is to 15
Kind target antibiotic is detected；The chromatographic column of liquid chromatogram selects ACQUITYBEH C18 model column 2.1 ×
100mm, 1.7 μm.
Liquid chromatogram separation parameter are as follows: 35 DEG C of chromatographic column column temperature；2 μ L of sample volume；10 DEG C of sample introduction room temperature；Flow rate of mobile phase
0.25mL·min-1；Mobile phase A is formic acid-aqueous solution that volumetric concentration is 0.5%, and water therein is Milli-Q ultrapure water；
Mobile phase B is acetonitrile；Gradient elution program: 0-2.2min, 16%B；2.2-2.5min 16%B-95% B；2.5-5.5min
95%B；5.5-6.0min, 95%B-16%B；6.0-10.0min, 16%B.
Mass Spectrometer Method condition are as follows: use electric spray ion source positive ion mode ESI+, triple level four bars mass analyzers are swept
Retouch mode: multiple-reaction monitoring pattern MRM is detected；Collision gas is high-purity argon gas；Desolvation temperature is 500 DEG C, desolventizing
Air velocity is 1000L/Hr, orifice potential 30V, capillary voltage 3.5kV, sweep time 0.1S.
Consider that different classes of antibiotic structure and property have very big difference, has different journeys in pretreatment process
The loss of degree, so, sulfamethoxazole-is selected respectively13C6As sulfa antibiotics (sulphadiazine, sulfamethyldiazine, sulphur
Amine dimethyl pyrimidine, sulphadimethoxine, sulfamethoxazole) internal standard compound, select Norfloxacin-D5As quinolone
The internal standard compound of class antibiotic (ciprofloxacin hydrochloride, Norfloxacin, Enrofloxacin) selects tetracycline-D6As Tetracyclines
The internal standard compound of antibiotic (aureomycin, terramycin, Doxycycline Hyclate) selects potassium penicillin G-D5It is anti-as itrofurans
The internal standard compound of raw element (furazolidone), selects trimethoprim-D3As the internal standard compound of trimethoprim, olaquindox-D is selected4As quinoline
The internal standard compound of ethyl alcohol selects Amoxicillin-13C6As the internal standard compound of beta-lactam antibiotic (Amoxicillin), due to internal standard
Object is selected appropriately, so that final detection result is more true and reliable.Consider that internal standard substance nature is more stable, in positive EFI
It responds under mist ionization source (ESI+) mode well and without obvious matrix interference.
The rate of recovery calculates:
Pre-treatment is carried out using the pre-treating method of step 1 to step 2, quantitative inspection is carried out using the method for step 3
It surveys, recovery of standard addition calculating is carried out to aquaculture water sample, calculated result is shown in Table 1.
The calculation formula of recovery of standard addition (RE%) are as follows:
Wherein, RE: recovery of standard addition, %；
C0: the concentration of hybrid standard liquid, ng/L；
C1: the detectable concentration of the water sample of hybrid standard liquid, ng/L are not added；
C2: the detectable concentration of the water sample of mixed standard solution, ng/L is added；
V0: the volume of hybrid standard liquid, L；
V1: constant volume, L machine before are not added on the water sample of hybrid standard liquid；
V2: constant volume before machine, L are added on the water sample of mixed standard solution.
It can be seen that the recovery of standard addition of 15 kinds of antibiotic is in 60%- in the water sample measured in the present inventive method
120%, recovery of standard addition has differences, and illustrates that there are matrix interferences；Recovery of standard addition is calculated by addition internal standard compound, and uses it
Inspection result is corrected, the influence of matrix interference bring can be reduced to a certain extent.
The recovery of standard addition of the measured concentration of antibiotic and different spiked levels in 1. water sample of table
Embodiment 3: practical aquaculture water sample antibiotic content measurement
The measurement of concentration of 15 kinds of antibiotic in two aquatic farms
The water sample for acquiring Shanghai City crab farming field, supporting shrimp first carries out specimen preprocessing using the method for step 1 of the present invention
Reason, then enrichment purification pre-treatment is carried out using the solid phase extraction method of step 2 of the present invention, later using step 3 of the present invention
High performance liquid chromatography tandem mass spectrum instrument tests and analyzes the actual concentrations of sample, investigates method of the invention to difference with this
The applicability of aquatic farm.
Specific detection process is as follows:
(1) water sample pre-processes
Each sample sets three in parallel, and the accurate 1000mL water sample that measures is in clean beaker respectively, through glass fiber filter
It is filtered to remove graininess suspended matter, then adjusts pH to 3 or so with dilute hydrochloric acid.
The sulfamethoxazole-of 100ng is added before Solid Phase Extraction into sample respectively13C6, trimethoprim-D3, benzyl penicillin
Sylvite-D5, Norfloxacin-D5, tetracycline-D6, olaquindox-D4With Amoxicillin-13C6The internal standard compound of this seven kinds instruction rate of recovery,
It is sufficiently mixed uniformly with water sample.That is: sulfamethoxazole-is selected respectively13C6As sulfa antibiotics (sulphadiazine, sulfalene
Yl pyrimidines, sulfamethazine, sulphadimethoxine, sulfamethoxazole) internal standard compound, select Norfloxacin-D5
As the internal standard compound of quinolone antibiotics (ciprofloxacin hydrochloride, Norfloxacin, Enrofloxacin), tetracycline-D is selected6Make
For the internal standard compound of tetracycline antibiotics (aureomycin, terramycin, Doxycycline Hyclate), potassium penicillin G-D is selected5As nitre
The internal standard compound of base furans antibiotic (furazolidone) selects trimethoprim-D3As the internal standard compound of trimethoprim, quinoline second is selected
Alcohol-D4As the internal standard compound of olaquindox, Amoxicillin-is selected13C6Internal standard as beta-lactam antibiotic (Amoxicillin)
Object.
(2) solid phase extraction concentration target antibiotic
5mL methyl tertiary butyl ether(MTBE), methanol, ultrapure water activated solid extraction column are successively used, flow control is in 4mL/min.In advance
Treated 1000mL water sample crosses column with the speed of 5mL/min, is successively with 6mL ultrapure water and isometric concentration after completion of the sample
5% methanol-water solution elutes solid-phase extraction column, vacuum drying with the speed of 4mL/min.Finally use the methanol (HPLC of 10mL
Grade methanol) elution, flow control is in 4mL/min, and eluent is collected into tool plug glass centrifuge tube, and eluent is under water bath condition
(40 DEG C of heating water bath) dries up under nitrogen stream effect, and the residue obtained methanol-water solution for being 70% with volumetric concentration is settled to
After 1mL, ultrasonic 2min, it is transferred in brown sample injection bottle after 0.22 μm of PTFE pin type filter filters, it is to be measured.
Wherein, the solid-phase extraction column that enrichment method sample uses is Waters company Oasis HLB pillar, and adsorbent is
The macroporous copolymer aggregated by a certain percentage by two kinds of monomers of lipophilicity divinylbenzene and hydrophily n-vinyl pyrrolidone,
It is the universal adsorbent for acidic, neutral and basic compounds.
(3) high performance liquid chromatography tandem mass spectrometry measures the content of 15 kinds of antibiotic in water body
Using internal standard method, on high performance liquid chromatography tandem mass spectrum instrument, it is enriched in quantitative detection brown sample injection bottle from water body
The 15 kinds of antibiotic arrived.15 kinds of antibiotic refer to that aquaculture is common or forbidden substance, including sulfamido, quinoline promise
Ketone, Tetracyclines, beta-lactam, itrofurans, trimethoprim, olaquindox these seventh types antibiotic；15 kinds of antibiotic point
It Wei not sulphadiazine, sulfamethyldiazine, sulphadimethoxine, trimethoprim, olaquindox, sulfamethoxazole, sulfanilamide (SN)
Diformazan pyrimidine, Amoxicillin, furazolidone, ciprofloxacin hydrochloride, Norfloxacin, Enrofloxacin, aureomycin, terramycin and salt
Sour Doxycycline.
Select the Waters of Waters, USTQ-Smicro high performance liquid chromatography tandem mass spectrum instrument is to 15
Kind target antibiotic is detected；The chromatographic column of liquid chromatogram selects ACQUITYBEH C18 column (2.1 ×
100mm, 1.7 μm).
Liquid chromatogram separation parameter are as follows: 35 DEG C of chromatographic column column temperature；2 μ L of sample volume；10 DEG C of sample introduction room temperature；Flow rate of mobile phase
0.25mL·min-1；Mobile phase A is formic acid-aqueous solution that volumetric concentration is 0.5%, and water therein is Milli-Q ultrapure water；
Mobile phase B is acetonitrile；Gradient elution program: 0-2.2min, 16%B；2.2-2.5min 16%B-95% B；2.5-5.5min
95%B；5.5-6.0min, 95%B-16%B；6.0-10.0min, 16%B.
Mass Spectrometer Method condition are as follows: use electric spray ion source positive ion mode ESI+, triple level four bars mass analyzers are swept
Retouch mode: multiple-reaction monitoring pattern MRM is detected；Collision gas is high-purity argon gas；Desolvation temperature is 500 DEG C, desolventizing
Air velocity is 1000L/Hr, orifice potential 30V, capillary voltage 3.5kV, sweep time 0.1S.
15 kinds of antibiotic contents are as shown in table 2 in two kinds of aquatic farms of actually detected gained.The experimental results showed that this hair
Bright method can be applied to the measurement of the antibiotic content of crab farming field and feeding shrimp, have high sensitivity, stability high and again
The good advantage of existing property.
The detection level of 15 kinds of antibiotic in 2 two kinds of aquaculture water samples of table
Note: " ND " expression is not detected.
Claims (10)
1. the side that a kind of Solid Phase Extraction pre-treatment combination LC-MS technology detects 15 kinds of antibiotic in aquaculture system simultaneously
Method, which is characterized in that sequentially include the following steps:
(1) pretreatment of water sample
Water sample is separately added into sulfamethoxazole-after membrane filtration removes suspended matter13C6, trimethoprim-D3, potassium penicillin G-
D5, Norfloxacin-D5, tetracycline-D6, olaquindox-D4With Amoxicillin-13C6The internal standard compound of this seven kinds instruction rate of recovery；It adjusts
Water sample pH；
(2) solid phase extraction concentration target antibiotic
Successively use methyl tertiary butyl ether(MTBE), methanol, ultrapure water activated solid extraction column；By processed water sample in step (1) after
The solid-phase extraction column of activation；Solid-phase extraction column is eluted with leacheate after the completion of enrichment and is dried in vacuo；Washing with certain volume again
Desolventizing elutes object, collects eluent and dries up under nitrogen flowing, residue obtained to re-dissolve, constant volume；It is shifted after filtering
It is to be measured into sample injection bottle；
(3) high performance liquid chromatography tandem mass spectrometry measures the content of seven 15 kinds of antibiotic of class in water body
Using internal standard method, on high performance liquid chromatography tandem mass spectrum instrument, interior 15 kinds be enriched to from water body of quantitative detection sample injection bottle
The content of antibiotic；15 kinds of antibiotic include sulfamido, quinolones, Tetracyclines, beta-lactam, nitrofuran
Class, trimethoprim, olaquindox these seventh types antibiotic；15 kinds of antibiotic are respectively two between sulphadiazine, sulfamethyldiazine, sulfanilamide (SN)
Methoxy pyrimidine, trimethoprim, olaquindox, sulfamethoxazole, sulfadimidine, Amoxicillin, furazolidone, Ciprofloxacin
Hydrochloride, Norfloxacin, Enrofloxacin, aureomycin, terramycin and Doxycycline Hyclate.
2. the method according to claim 1, wherein the sulfalene for being separately added into 100ng/L is disliked in step (1)
Azoles-13C6, trimethoprim-D3, potassium penicillin G-D5, Norfloxacin-D5, tetracycline-D6, olaquindox-D4With Amoxicillin-13C6
This seven kinds of internal standard compounds indicate the rate of recovery.
3. the method according to claim 1, wherein water sample is filtered to remove through glass fiber filter in step (1)
After graininess suspended matter, the internal standard compound of the instruction rate of recovery is first added, then adjust pH value to 3 ± 0.1 with dilute hydrochloric acid.
4. according to the method described in claim 3, it is characterized in that, water sample filters glass fiber filter used in step (1)
For 0.7 μm of aperture Whatman GF/F Series glass fiber filter membrane.
5. method according to claim 1,2 or 3, which is characterized in that in step (2), solid-phase extraction column used is
Waters company Oasis HLB pillar, adsorbent are by lipophilicity divinylbenzene and hydrophily n-vinyl pyrrolidone two
The macroporous copolymer that kind monomer aggregates by a certain percentage, is the universal adsorbent for acidic, neutral and basic compounds.
6. method according to claim 1,2 or 3, which is characterized in that in step (2), used in activated solid extraction column
Methyl tertiary butyl ether(MTBE), methanol and water consumption are respectively 1 times for extracting column volume, and the flow control of activated solid extraction column is in 1-4mL/
min；Water sample crosses column flow rate control in 3-5mL/min；It elutes pure water used in extraction column and low concentration methanol aqueous solution is respectively extraction
Take 1-2 times of column volume；It is drained after elution, time 5-10min；Eluting eluting solvent used in solid-phase extraction column is methanol,
The amount of methanol eluting solvent is 2 times for extracting column volume, and eluent flow rate is controlled in 4-5mL/min.
7. method according to claim 1,2 or 3, which is characterized in that in step (2), elute low concentration methanol used
Methanol-water solution for volumetric concentration less than 5%；Elution methanol used is HPLC grades of methanol；Methanol used in constant volume residue
It is the methanol-water solution that volumetric concentration is 70%；Eluent nitrogen under the conditions of 40 DEG C of heating water bath is blown to close dry；Use methanol constant volume
Afterwards, ultrasonic 2min, then filtered with pin type filter；Pin type filter selects polytetrafluoroethylene (PTFE) pin type filter of the aperture less than 0.22 μm.
8. method according to claim 1,2 or 3, which is characterized in that in step (3), select Waters, US
WatersTQ-Smicro high performance liquid chromatography tandem mass spectrum instrument detects 15 kinds of target antibiotic；Liquid chromatogram
Chromatographic column select ACQUITYBEH C18 model column 2.1 × 100mm, 1.7 μm.
9. method according to claim 1,2 or 3, which is characterized in that in step (3), liquid chromatogram separation parameter are as follows: color
35 DEG C of column column temperature of spectrum；2 μ L of sample volume；10 DEG C of sample introduction room temperature；Flow rate of mobile phase 0.25mLmin-1；Mobile phase A is that volume is dense
Formic acid-aqueous solution that degree is 0.5%, water therein are Milli-Q ultrapure water；Mobile phase B is acetonitrile；Gradient elution program: 0-
2.2min, 16%B；2.2-2.5min, 16%B-95%B；2.5-5.5min, 95%B；5.5-6.0min, 95%B-16%B；
6.0-10.0min, 16%B.
10. method according to claim 1,2 or 3, which is characterized in that in step (3), Mass Spectrometer Method condition are as follows: use
Electric spray ion source positive ion mode ESI+, triple level four bars mass analyzers, scanning mode: multiple-reaction monitoring pattern MRM into
Row detection；Collision gas is high-purity argon gas；Desolvation temperature is 500 DEG C, and Desolvention gas velocity degree is 1000L/Hr, orifice potential
For 30V, capillary voltage 3.5kV, sweep time 0.1S.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110320301A (en) * | 2019-07-22 | 2019-10-11 | 东莞正大康地饲料有限公司 | The detection method of olaquindox in a kind of premix containing Organic Chromium |
| CN110514762A (en) * | 2019-08-30 | 2019-11-29 | 福建佳泛美实验室系统工程有限公司 | A kind of method that the dedicated column solid phase extraction of water sample with bulk mass-Liquid Chromatography-Tandem Mass Spectrometry measures antibiotic in water |
| CN111505149A (en) * | 2020-04-30 | 2020-08-07 | 华东理工大学 | Method for detecting sulfanilamide and β -lactam antibiotics in livestock and poultry breeding wastewater |
| CN111751482A (en) * | 2020-06-05 | 2020-10-09 | 哈尔滨工业大学 | Method for synchronously detecting residual quantity of various antibiotics in pig-raising wastewater |
| CN114720572A (en) * | 2020-12-22 | 2022-07-08 | 上海市环境科学研究院 | Method for detecting content of 15 antibiotics in fish meat |
| CN116879462A (en) * | 2023-09-04 | 2023-10-13 | 北京师范大学 | Method, device and system for analyzing agricultural chemicals and metabolites thereof in serum |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105758955A (en) * | 2016-02-26 | 2016-07-13 | 清华大学 | Method for determining antibiotics in water environment |
| CN107219310A (en) * | 2017-05-16 | 2017-09-29 | 河南省兽药饲料监察所 | The screening method of 110 kinds of medicines in a kind of feed |
| CN107490649A (en) * | 2017-09-15 | 2017-12-19 | 河南省兽药饲料监察所 | The screening method of 62 kinds of antibacterials in a kind of farm animal excrement |
| CN107543877A (en) * | 2017-06-09 | 2018-01-05 | 上海市环境科学研究院 | A kind of method that SPE liquid chromatography tandem mass spectrometry determines six 24 kinds of antibiotic of class in water body simultaneously |
| CN107561205A (en) * | 2017-08-24 | 2018-01-09 | 中国水产科学研究院南海水产研究所 | A kind of method of 45 kinds of veterinary drugs in LC-MS detection pond deposit |
| CN107561187A (en) * | 2017-09-08 | 2018-01-09 | 中国科学院生态环境研究中心 | A kind of method of Multiple Classes of Antibiotics in synchronous detection polluted-water |
| CN107907620A (en) * | 2017-11-22 | 2018-04-13 | 上海市环境科学研究院 | The method that ultrasonic extraction Solid Phase Extraction pre-treatment combination LC-MS technology detects six 24 kinds of antibiotic of class in feces of livestock and poultry at the same time |
| CN107941928A (en) * | 2017-10-24 | 2018-04-20 | 东莞市环境监测中心站 | A kind of method of antibiotic in liquid chromatography mass Simultaneous Determination environment water |
| CN108318613A (en) * | 2018-01-11 | 2018-07-24 | 四川国测检测技术有限公司 | The detection method of antibiotic in a kind of environmental sample |
-
2018
- 2018-12-07 CN CN201811495037.0A patent/CN109342624A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105758955A (en) * | 2016-02-26 | 2016-07-13 | 清华大学 | Method for determining antibiotics in water environment |
| CN107219310A (en) * | 2017-05-16 | 2017-09-29 | 河南省兽药饲料监察所 | The screening method of 110 kinds of medicines in a kind of feed |
| CN107543877A (en) * | 2017-06-09 | 2018-01-05 | 上海市环境科学研究院 | A kind of method that SPE liquid chromatography tandem mass spectrometry determines six 24 kinds of antibiotic of class in water body simultaneously |
| CN107561205A (en) * | 2017-08-24 | 2018-01-09 | 中国水产科学研究院南海水产研究所 | A kind of method of 45 kinds of veterinary drugs in LC-MS detection pond deposit |
| CN107561187A (en) * | 2017-09-08 | 2018-01-09 | 中国科学院生态环境研究中心 | A kind of method of Multiple Classes of Antibiotics in synchronous detection polluted-water |
| CN107490649A (en) * | 2017-09-15 | 2017-12-19 | 河南省兽药饲料监察所 | The screening method of 62 kinds of antibacterials in a kind of farm animal excrement |
| CN107941928A (en) * | 2017-10-24 | 2018-04-20 | 东莞市环境监测中心站 | A kind of method of antibiotic in liquid chromatography mass Simultaneous Determination environment water |
| CN107907620A (en) * | 2017-11-22 | 2018-04-13 | 上海市环境科学研究院 | The method that ultrasonic extraction Solid Phase Extraction pre-treatment combination LC-MS technology detects six 24 kinds of antibiotic of class in feces of livestock and poultry at the same time |
| CN108318613A (en) * | 2018-01-11 | 2018-07-24 | 四川国测检测技术有限公司 | The detection method of antibiotic in a kind of environmental sample |
Non-Patent Citations (6)
| Title |
|---|
| LI-JUN ZHOU ET AL: "Simultaneous determination of human and veterinary antibiotics in various environmental matrices by rapid resolution liquid chromatography–electrospray ionization tandem mass spectrometry", 《JOURNAL OF CHROMATOGRAPHY A》 * |
| S. BORRÀS ET AL: "Multiclass method for antimicrobial analysis in animal feeds by liquid chromatography–tandem mass spectrometry", 《ANAL BIOANAL CHEM》 * |
| YI LUO ET AL: "Occurrence and Transport of Tetracycline, Sulfonamide, Quinolone,and Macrolide Antibiotics in the Haihe River Basin, China", 《ENVIRONMENTAL SCIENCE & TECHNOLOGY》 * |
| 何欣 等: "基于高效液相色谱―四级杆―飞行时间质谱技术筛查渔业水域环境中抗生素残留方法的建立与应用", 《水产学报》 * |
| 吕佳 等: "饮用水中35种抗生素的超高效液相色谱-串联质谱测定法", 《环境与健康杂志》 * |
| 王军淋 等: "固相萃取富集－超高效液相色谱－串联质谱法同时测定源水和自来水中的40种多类别抗生素", 《中国卫生检验杂志》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110320301A (en) * | 2019-07-22 | 2019-10-11 | 东莞正大康地饲料有限公司 | The detection method of olaquindox in a kind of premix containing Organic Chromium |
| CN110514762A (en) * | 2019-08-30 | 2019-11-29 | 福建佳泛美实验室系统工程有限公司 | A kind of method that the dedicated column solid phase extraction of water sample with bulk mass-Liquid Chromatography-Tandem Mass Spectrometry measures antibiotic in water |
| CN111505149A (en) * | 2020-04-30 | 2020-08-07 | 华东理工大学 | Method for detecting sulfanilamide and β -lactam antibiotics in livestock and poultry breeding wastewater |
| CN111751482A (en) * | 2020-06-05 | 2020-10-09 | 哈尔滨工业大学 | Method for synchronously detecting residual quantity of various antibiotics in pig-raising wastewater |
| CN111751482B (en) * | 2020-06-05 | 2022-12-23 | 哈尔滨工业大学 | Method for synchronously detecting residual quantity of various antibiotics in pig-raising wastewater |
| CN114720572A (en) * | 2020-12-22 | 2022-07-08 | 上海市环境科学研究院 | Method for detecting content of 15 antibiotics in fish meat |
| CN114720572B (en) * | 2020-12-22 | 2023-11-24 | 上海市环境科学研究院 | Method for detecting 15 antibiotics content in fish meat |
| CN116879462A (en) * | 2023-09-04 | 2023-10-13 | 北京师范大学 | Method, device and system for analyzing agricultural chemicals and metabolites thereof in serum |
| CN116879462B (en) * | 2023-09-04 | 2023-11-17 | 北京师范大学 | Method, device and system for analyzing agricultural chemicals and metabolites thereof in serum |
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